C7-Fluoro Substituted Tetracycline Compounds

ABSTRACT

The present invention is directed to a compound represented by Structural Formula (A): 
     
       
         
         
             
             
         
       
     
     or a pharmaceutically acceptable salt thereof. The variables for Structural Formula (A) are defined herein. Also described is a pharmaceutical composition comprising the compound of Structural Formula (A) and its therapeutic use.

RELATED APPLICATION

This application is a continuation of U.S. application Ser. No.14/532,882, filed Nov. 4, 2014, which is a continuation of U.S.application Ser. No. 12/462,795, filed Aug. 7, 2009, now U.S. Pat. No.8,906,887, which claims the benefit of U.S. Provisional Application No.61/188,307, filed on Aug. 8, 2008. The entire teachings of the aboveapplications are incorporated herein by reference.

BACKGROUND OF THE INVENTION

The tetracyclines are broad spectrum anti-microbial agents that arewidely used in human and veterinary medicine. The total production oftetracyclines by fermentation or semi-synthesis is measured in thethousands of metric tons per year.

The widespread use of tetracyclines for therapeutic purposes has led tothe emergence of resistance to these antibiotics, even among highlysusceptible bacterial species. Therefore, there is need for newtetracycline analogs with improved antibacterial activities andefficacies against other tetracycline responsive diseases or disorders.

SUMMARY OF THE INVENTION

The present invention is directed to a compound represented byStructural Formula (A):

or a pharmaceutically acceptable salt thereof, wherein:

X is selected from hydrogen, —(C₁-C₇)alkyl, carbocyclyl, aryl andheteroaryl;

Y is selected from hydrogen, —(C₁-C₇)alkyl, carbocyclyl,—(C₁-C₄)alkylene-N(R^(A))(R^(B)),—(C₁-C₄)alkylene-N(R^(F))—C(O)—[C(R^(D))(R^(E))]₀₋₄—N(R^(A))(R^(B)),—CH═N—OR^(A), —N(R^(A))(R^(B)),—N(R^(F))—C(O)—[C(R^(D))(R^(E))]₁₋₄—N(R^(A))(R^(B)),—N(R^(F))—C(O)—N(R^(A))(R^(B)), —N(R^(F))—C(O)—(C₁-C₆)alkyl,—N(R^(F))—C(O)-heterocyclyl, —N(R^(F))—C(O)-heteroaryl,—N(R^(F))—C(O)-carbocyclyl, —N(R^(F))—C(O)-aryl,—N(R^(F))—S(O)_(m)—(C₁-C₄)alkylene-N(R^(A))(R^(B)),—N(R^(F))—S(O)_(m)—(C₁-C₄)alkylene-carbocyclyl, and—N(R^(F))—S(O)_(m)—(C₁-C₄)alkylene-aryl;

at least one of X and Y is not hydrogen;

each R^(A) and R^(B) are independently selected from hydrogen,(C₁-C₇)alkyl, —O—(C₁-C₇)alkyl, —(C₀-C₆)alkylene-carbocyclyl,—(C₀-C₆)alkylene-aryl, —(C₀-C₆)alkylene-heterocyclyl,—(C₀-C₆)alkylene-heteroaryl, —(C₁-C₆)alkylene-O-carbocyclyl,—(C₁-C₆)alkylene-O-aryl, —(C₁-C₆)alkylene-O-heterocyclyl,—(C₁-C₆)alkylene-O-heteroaryl, —S(O)_(m)—(C₁-C₆)alkyl,—(C₀-C₄)alkylene-S(O)_(m)-carbocyclyl, —(C₀-C₄)alkylene-S(O)_(m)-aryl,—(C₀-C₄)alkylene-S(O)_(m)-heterocyclyl and—(C₀-C₄)alkylene-S(O)_(m)-heteroaryl; or

R^(A) and R^(B) taken together with the nitrogen atom to which they arebound form a heterocyclyl or heteroaryl, wherein the heterocycle orheteroaryl optionally comprises 1 to 4 additional heteroatomsindependently selected from N, S and O;

each R^(D) and each R^(E) is independently selected from hydrogen,(C₁-C₆)alkyl, carbocyclyl, aryl, heterocyclyl, heteroaryl, or anaturally occurring amino acid side chain moiety, or

R^(D) and R^(E) taken together with the carbon atom to which they arebound form a 3-7 membered carbocyclyl, or a 4-7 membered heterocyclyl,wherein the heterocyclyl formed by R^(D) and R^(E) optionally comprisesone to two additional heteroatoms independently selected from N, S andO;

R^(F) is selected from hydrogen, (C₁-C₇)alkyl, carbocyclyl, aryl andheteroaryl; and

m is 1 or 2, wherein:

each carbocyclyl, aryl, heterocyclyl or heteroaryl is optionally andindependently substituted with one or more substituents independentlyselected from halo, —(C₁-C₄)alkyl, —OH, ═O, —O—(C₁-C₄)alkyl,—(C₁-C₄)alkyl-O—(C₁-C₄)alkyl, halo-substituted —(C₁-C₄)alkyl,halo-substituted —O—(C₁-C₄)alkyl, —C(O)—(C₁-C₄)alkyl,—C(O)-(fluoro-substituted-(C₁-C₄)alkyl), —S(O)_(m)—(C₁-C₄)alkyl,—N(R^(G))(R^(G)), and CN;

each alkyl in the group represented by R^(A), R^(B), R^(D) and R^(E) isoptionally and independently substituted with one or more substituentsindependently selected from halo, —(C₁-C₄)alkyl, —OH, —O—(C₁-C₇)alkyl,—(C₁-C₄)alkyl-O—(C₁-C₄)alkyl, fluoro-substituted-(C₁-C₄)alkyl,—S(O)_(m)—(C₁-C₄)alkyl, and —N(R^(G))(R^(G)), wherein

each R^(G) is hydrogen or (C₁-C₄)alkyl, wherein each alkyl in the grouprepresented by R^(G) is optionally and independently substituted withone or more substituents independently selected from —(C₁-C₄)alkyl,(C₃-C₆)cycloalkyl, halo, —OH, —O—(C₁-C₄)alkyl, and(C₁-C₄)alkyl-O—(C₁-C₄)alkyl.

Another embodiment of the present invention is directed to a compoundrepresented by Structural Formula (II)

or a pharmaceutically acceptable salt thereof, wherein:

R¹ and R² are each independently selected from hydrogen, (C₁-C₇)alkyl,(C₃-C₆)cycloalkyl(C₁-C₄)alkyl, (C₁-C₇)alkoxy(C₁-C₄)alkyl,(C₃-C₆)cycloalkoxy(C₁-C₄)alkyl, (C₃-C₆)cycloalkyl, aryl,aryl(C₁-C₄)alkyl, aryloxy(C₁-C₄)alkyl, arylthio(C₁-C₄)alkyl,arylsufinyl(C₁-C₄)alkyl, arylsulfonyl(C₁-C₄)alkyl, and —O—(C₁-C₇)alkyl,or

R¹ and R² taken together with the nitrogen atom to which they are bondedform a monocyclic or bicyclic heteroaryl, or a monocyclic, fusedbicyclic, bridged bicyclic or spiro bicyclic heterocycle, wherein theheteroaryl or heterocycle optionally contains one or two additionalheteroatoms independently selected from N, O and S; and

wherein each alkyl, cycloalkyl, alkoxy and cycloalkoxy moiety in thegroups represented by R¹ and R² and each heterocycle represented byNR¹R² taken together is optionally substituted with one or moresubstituents independently selected from the group consisting of(C₁-C₄)alkyl, halo, —OH, (C₁-C₄)alkoxy, (C₁-C₄)alkylthio,(C₁-C₄)alkylsulfinyl, (C₁-C₄)alkylsulfonyl, (C₁-C₄)alkoxy(C₁-C₄)alkyl,and —N(R³)(R⁴); and

each aryl, aryloxy, arylthio, arylsufinyl and arylsulfonyl moiety in thegroups represented by R¹ and R² and each heteroaryl represented by NR¹R²taken together is optionally substituted with one or more substituentsindependently selected from the group consisting of (C₁-C₄)alkyl, halo,—OH, (C₁-C₄)alkoxy, —S—(C₁-C₄)alkyl, —S(O)(C₁-C₄)alkyl,—S(O)₂(C₁-C₄)alkyl, (C₁-C₄)alkoxy(C₁-C₄)alkyl, —N(R³)(R⁴); —CN,halo(C₁-C₄)alkyl, and halo(C₁-C₄)alkoxy, and

R³ and R⁴ are each independently selected from the group consisting of—H and (C₁-C₄)alkyl, wherein the (C₁-C₄)alkyl represented by R³ and R⁴is optionally substituted with one or more substituents independentlyselected from the group consisting of (C₁-C₄)alkyl, halo, —OH,(C₁-C₄)alkoxy, and (C₁-C₄)alkoxy(C₁-C₄)alkyl. Values for X and R^(F) areas described above for Structural Formula (A).

Another embodiment of the present invention is directed to a compoundrepresented by Structural Formula (I):

or a pharmaceutically acceptable salt thereof, wherein:

R¹ and R² are each independently selected from hydrogen, (C₁-C₇)alkyl,(C₃-C₆)cycloalkyl(C₁-C₄)alkyl, (C₁-C₇)alkoxy(C₁-C₄)alkyl,(C₃-C₆)cycloalkoxy(C₁-C₄)alkyl, (C₃-C₆)cycloalkyl, aryl,aryl(C₁-C₄)alkyl, aryloxy(C₁-C₄)alkyl, arylthio(C₁-C₄)alkyl,arylsulfinyl(C₁-C₄)alkyl, and arylsulfonyl(C₁-C₄)alkyl, or

R¹ and R² taken together with the nitrogen atom to which they are bondedform a monocyclic or bicyclic heteroaryl, or a monocyclic, fusedbicyclic, bridged bicyclic or spiro bicyclic heterocycle, wherein theheteroaryl or heterocycle optionally contains one additional heteroatomindependently selected from N, O and S; and the remainder of thevariables are as described above for Structural Formula (II).

Another embodiment of the present invention is directed to apharmaceutical composition comprising a pharmaceutically acceptablecarrier or diluent and a compound represented by Structural Formula (A),(II) or (I) or a pharmaceutically acceptable salt thereof. Thepharmaceutical composition is used in therapy, such as treating aninfection in a subject.

Another embodiment of the present invention is a method of treating aninfection in a subject comprising administering to the subject aneffective amount of a compound represented by Structural Formula (A),(II) or (I) or a pharmaceutically acceptable salt thereof.

Another embodiment of the present invention is a method of preventing aninfection in a subject comprising administering to the subject aneffective amount of a compound represented by Structural Formula (A),(II) or (I) or a pharmaceutically acceptable salt thereof.

Another embodiment of the present invention is the use of a compoundrepresented by Structural Formula (A), (II) or (I) or a pharmaceuticallyacceptable salt thereof for the manufacture of a medicament for treatingan infection in a subject.

Another embodiment of the present invention is the use of a compoundrepresented by Structural Formula (A), (II) or (I) or a pharmaceuticallyacceptable salt thereof for the manufacture of a medicament forpreventing an infection in a subject.

Another embodiment of the present invention is the use of a compoundrepresented by Structural Formula (A), (II) or (I) or a pharmaceuticallyacceptable salt thereof for therapy, such as treating or preventing aninfection in a subject.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed to a compound represented byStructural Formula (A) or a pharmaceutically acceptable salt thereof.Values and alternative values for the variables in Structural Formula(A) are defined as the following:

X is selected from hydrogen, —(C₁-C₇)alkyl, carbocyclyl, aryl andheteroaryl. In one embodiment, X is hydrogen. In an alternativeembodiment, X is —(C₁-C₇)alkyl. Alternatively, X is —(C₁-C₄)alkyl. Inanother embodiment, X is carbocyclyl. In another alternative embodiment,X is aryl or heteroaryl. In another alternative embodiment, X is phenyl.

Y is selected from hydrogen, —(C₁-C₇)alkyl,

carbocyclyl, —(C₁-C₄)alkylene-N(R^(A))(R^(B)),—(C₁-C₄)alkylene-N(R^(F))—C(O)—[C(R^(D))(R^(E))]₀₋₄—N(R^(A))(R^(B)),—CH═N—OR^(A), —N(R^(A))(R^(B)),—N(R^(F))—C(O)—[C(R^(D))(R^(E))]₁₋₄—N(R^(A))(R^(B)),—N(R^(F))—C(O)—N(R^(A))(R^(B)), —N(R^(F))—C(O)—(C₁-C₆)alkyl,—N(R^(F))—C(O)-heterocyclyl, —N(R^(F))—C(O)-heteroaryl,—N(R^(F))—C(O)-carbocyclyl, —N(R^(F))—C(O)-aryl,—N(R^(F))—S(O)_(m)—(C₁-C₄)alkylene-N(R^(A))(R^(B)),—N(R^(F))—S(O)_(m)—(C₁-C₄)alkylene-carbocyclyl,and —N(R^(F))—S(O)_(m)—(C₁-C₄)alkylene-aryl, provided at least one of Xand Y is not hydrogen. In one embodiment, Y is selected from hydrogen,—(C₁-C₇)alkyl, —(C₁-C₄)alkylene-N(R^(A))(R^(B)),—(C₁-C₄)alkylene-N(R^(F))—C(O)—[C(R^(D))(R^(E))]₀₋₄—N(R^(A))(R^(B)),—CH═N—OR^(A), —N(R^(A))(R^(B)),—N(R^(F))—C(O)—[C(R^(D))(R^(E))]₁₋₄—N(R^(A))(R^(B)),—N(R^(F))—C(O)—N(R^(A))(R^(B)), —N(R^(F))—C(O)—(C₁-C₆)alkyl,—N(R^(F))—C(O)-heterocyclyl, —N(R^(F))—C(O)-heteroaryl,—N(R^(F))—C(O)-carbocyclyl, —N(R^(F))—C(O)-aryl,—N(R^(F))—S(O)_(m)—(C₁-C₄)alkylene-N(R^(A))(R^(B)),—N(R^(F))—S(O)_(m)—(C₁-C₄)alkylene-carbocyclyl,and —N(R^(F))—S(O)_(m)—(C₁-C₄)alkylene-aryl, provided at least one of Xand Y is not hydrogen. In one embodiment, Y is selected from hydrogen,—(C₁-C₇)alkyl, —(C₁-C₄)alkylene-N(R^(A))(R^(B)),—(C₁-C₄)alkylene-N(R^(F))—C(O)—[C(R^(D))(R^(E))]₀₋₄—N(R^(A))(R^(B)),—CH═N—OR^(A), —N(R^(A))(R^(B)),—N(R^(F′))—C(O)—[C(R^(D))(R^(E))]₁₋₄—N(R^(A))(R^(B)),—NH—C(O)—C(R^(D′))(R^(E))—N(R^(A))(R^(B)),—N(R^(F))—C(O)—N(R^(A))(R^(B)), —N(R^(F))—C(O)—(C₁-C₆)alkyl, —N(R^(F))—C(O)-heterocyclyl, —N(R^(F))—C(O)-heteroaryl, —N(R^(F))—C(O)-carbocyclyl,—N(R^(F))—C(O)-aryl —N(R^(F))—S(O)_(m)—(C₁-C₄)alkylene-N(R^(A))(R^(B)),—N(R^(F))—S(O)_(m)—(C₁-C₄)alkylene-carbocyclyl, and—N(R^(F))—S(O)_(m)—(C₁-C₄)alkylene-aryl. In another embodiment, Y isselected from hydrogen, —(C₁-C₇)alkyl, —(C₁-C₄)alkylene-N(R^(A))(R^(B)),—(C₁-C₄)alkylene-NH—C(O)—(CH₂)₀₋₁—N(R^(A))(R^(B)), —N(R^(A))(R^(B)),—NH—C(O)-carbocyclyl, —NH—C(O)-aryl, —NH—C(O)-heterocyclyl,—NH—C(O)-heteroaryl, —NH—C(O)—N(R^(A))(R^(A)),—N(R^(F′))—C(O)—CH₂—N(R^(A))(R^(B)),—NH—C(O)—C(R^(D′))(R^(E))—N(R^(A))(R^(B)) and—NH—S(O)_(m)—(C₁-C₄)alkylene-N(R^(A))(R^(B)). Alternatively, the—(C₁-C₇)alkyl represented by Y described above is a —(C₁-C₄)alkyl. Inyet another embodiment, Y is selectedfrom —N(R^(A))(R^(B)), —N(H)—C(O)-carbocyclyl, —N(H)—C(O)-aryl,—N(H)—C(O)-heterocycle, and —N(H)—C(O)-heteroaryl. Alternatively, Y is—NH—C(O)—CH₂—N(R^(A))(R^(B)). More specifically, R^(A) and R^(B) in—NH—C(O)—CH₂—N(R^(A))(R^(B)) are R¹ and R², respectively.

Each R^(A) and R^(B) are independently selected from hydrogen,(C₁-C₇)alkyl, —O—(C₁-C₇)alkyl, —(C₀-C₆)alkylene-carbocyclyl,—(C₀-C₆)alkylene-aryl, —(C₀-C₆)alkylene-heterocyclyl,—(C₀-C₆)alkylene-heteroaryl, —(C₁-C₆)alkylene-O-carbocyclyl,—(C₁-C₆)alkylene-O-aryl, —(C₁-C₆)alkylene-O-heterocyclyl,—(C₁-C₆)alkylene-O-heteroaryl, —S(O)_(m)—(C₁-C₆)alkyl,—(C₀-C₄)alkylene-S(O)_(m)-carbocyclyl, —(C₀-C₄)alkylene-S(O)_(m)-aryl,—(C₀-C₄)alkylene-S(O)_(m)-heterocyclyl and—(C₀-C₄)alkylene-S(O)_(m)-heteroaryl; or R^(A) and R^(B) taken togetherwith the nitrogen atom to which they are bound form a heterocyclyl orheteroaryl, wherein the heterocycle or heteroaryl optionally comprises 1to 4 additional heteroatoms independently selected from N, S and O. Inone embodiment, each R^(A) is independently selected from hydrogen andmethyl; R^(B) is selected from hydrogen,

(C₁-C₇)alkyl, —(C₀-C₆)alkylene-carbocyclyl, —(C₀-C₆)alkylene-aryl,—(C₀-C₆)alkylene-heteroaryl, —S(O)_(m)—(C₁-C₆)alkyl,—(C₀-C₄)alkylene-S(O)_(m)-carbocyclyl, —(C₀-C₄)alkylene-S(O)_(m)-aryl,—(C₀-C₄)alkylene-S(O)_(m)-heterocycle and—(C₀-C₄)alkylene-S(O)_(m)-heteroaryl; or R^(A) and R^(B) taken togetherwith the nitrogen atom to form a heterocycle, wherein the heterocycle isoptionally substituted with ═O and —N(R^(G))(R^(G)). In anotherembodiment, R^(A) is hydrogen; and R^(B) is selected from (C₁-C₄)alkyl,and —S(O)₂—CH₃; or R^(A) and R^(B) taken together to form 4-7 memberedheterocyclic ring.

Each R^(D) and each R^(E) is independently selected from hydrogen,(C₁-C₆)alkyl, carbocyclyl, aryl, heterocyclyl, heteroaryl, or anaturally occurring amino acid side chain moiety, or R^(D) and R^(E)taken together with the carbon atom to which they are bound form a 3-7membered carbocyclyl, or a 4-7 membered heterocyclyl, wherein theheterocyclyl formed by R^(D) and R^(E) optionally comprises one to twoadditional heteroatoms independently selected from N, S and O. In oneembodiment, R^(D) and R^(E) are both —H.

R^(F) is selected from hydrogen, (C₁-C₇)alkyl, carbocyclyl, aryl andheteroaryl. In one embodiment, R^(F) is hydrogen. In another embodiment,R is selected from hydrogen, (C₁-C₇)alkyl, aryl and heteroaryl. Inanother embodiment, R^(F) is selected from hydrogen, (C₁-C₇)alkyl andphenyl. In another embodiment, R^(F) is selected from hydrogen,(C₁-C₄)alkyl and phenyl

R^(D′) is selected from (C₁-C₆)alkyl, carbocyclyl, aryl, heteroaryl,heterocyclyl, and a naturally occurring amino acid side chain moiety, orR^(D′) and R^(E) taken together with the carbon atom to which they arebound form a 3-7 membered carbocyclyl, or a 4-7 membered heterocyclyl,wherein the heterocyclyl formed by R^(D′) and R^(E) optionally comprisesone to two additional heteroatoms independently selected from N, S andO. In one embodiment, R^(D′) and R^(E) taken together with the carbonatom to which they are bound form a (C₃-C₇)cycloalkyl.

m is 1 or 2. In one embodiment, m is 2.

Each carbocyclyl, aryl, heterocyclyl or heteroaryl described above(e.g., in the groups represented by Y, R^(A), R^(B), R^(D), R^(D′) andR^(E)) is optionally and independently substituted with one or moresubstituents independently selected from

halo, —(C₁-C₄)alkyl, —OH, ═O, —O—(C₁-C₄)alkyl,—(C₁-C₄)alkyl-O—(C₁-C₄)alkyl, halo-substituted —(C₁-C₄)alkyl,halo-substituted —O—(C₁-C₄)alkyl, —C(O)—(C₁-C₄)alkyl,—C(O)-(fluoro-substituted-(C₁-C₄)alkyl), —S(O)_(m)—(C₁-C₄)alkyl,—N(R^(G))(R^(G)), and CN. In one embodiment, each carbocyclyl,heterocyclyl or heteroaryl is optionally and independently substitutedwith one or more substituents independently selected from halo,—(C₁-C₄)alkyl,halo-substituted —(C₁-C₄)alkyl, —O—(C₁-C₄)alkyl, and —N(R^(G))(R^(G)).In another embodiment, each carbocyclyl, aryl, heteroaryl, orheterocycle is optionally and independently substituted with one or moresubstituents independently selected from —CH₃, fluoro, and —N(CH₃)₂.

Each alkyl described above (e.g., in the groups represented by Y, R^(A),R^(B), R^(D), R^(D′), R^(E), R^(F) and R^(F′)) is optionally andindependently substituted with one or more substituents independentlyselected from

halo, —(C₁-C₄)alkyl, —OH, —O—(C₁-C₇)alkyl, —(C₁-C₄)alkyl-O—(C₁-C₄)alkyl,fluoro-substituted-(C₁-C₄)alkyl, —S(O)_(m)—(C₁-C₄)alkyl, and—N(R^(G))(R^(G)). In one embodiment, each alkyl group (e.g., in thegroup represented by Y or R^(B)) is optionally and independentlysubstituted with one or more substituents independently selected fromhalo, —OH, and —N(R^(G))(R^(G)).

Each R^(G) is hydrogen or (C₁-C₄)alkyl, wherein each alkyl in the grouprepresented by R^(G) is optionally and independently substituted withone or more substituents independently selected from —(C₁-C₄)alkyl,(C₃-C₆)cycloalkyl, halo, —OH, —O—(C₁-C₄)alkyl, and(C₁-C₄)alkyl-O—(C₁-C₄)alkyl. In one embodiment, each alkyl in the grouprepresented by R^(G) is optionally and independently substituted with(C₃-C₆)cycloalkyl.

As used herein, when R^(A) and R^(B) taken together with the nitrogenatom to which they are bound form a a heterocyclyl or heteroaryl, theheterocyclyl or heteroaryl represented by —NR^(A)R^(B) can include aring system that has a heteroatom adjacent to the nitrogen atom to whichR^(A) and R^(B) are bound. For example, —NR^(A)R^(B) can be, but is notlimited to, the following ring systems:

Similarly, when R^(D) and R^(E) or R^(D′) and R^(E) taken together withthe carbon atom to which they are bound form a heterocyclyl, theheterocyclyl can include a ring system that has a heteroatom adjacent tothe carbon atom to which R^(D) and R^(E) or R^(D′) and R^(E) are bound.

The present invention is directed to a compound represented byStructural Formula (I) or (II) or a pharmaceutically acceptable saltthereof. Values and alternative values for the variables in StructuralFormula (I) or (II) are defined as the following:

R¹ is selected from hydrogen, (C₁-C₇)alkyl,(C₃-C₆)cycloalkyl(C₁-C₄)alkyl, (C₁-C₇)alkoxy(C₁-C₄)alkyl,(C₃-C₆)cycloalkoxy(C₁-C₄)alkyl, (C₃-C₆)cycloalkyl, aryl,aryl(C₁-C₄)alkyl, aryloxy(C₁-C₄)alkyl, arylthio(C₁-C₄)alkyl,arylsulfinyl(C₁-C₄)alkyl, arylsulfonyl(C₁-C₄)alkyl and —O—(C₁-C₇)alkyl.Each alkyl, cycloalkyl, alkoxy, cycloalkoxy, aryl, aryloxy, arylthio,arylsulfinyl and arylsulfonyl moiety in the groups represented by R¹ canbe optionally substituted with one or more independently selectedsubstituents defined above for Structural Formula (I). Alternatively, R¹is selected from hydrogen, (C₁-C₇)alkyl, (C₃-C₆)cycloalkyl(C₁-C₄)alkyl,(C₁-C₇)alkoxy(C₁-C₄)alkyl, (C₃-C₆)cycloalkoxy(C₁-C₄)alkyl,(C₃-C₆)cycloalkyl, aryl, aryl(C₁-C₄)alkyl, aryloxy(C₁-C₄)alkyl,arylthio(C₁-C₄)alkyl, arylsulfinyl(C₁-C₄)alkyl andarylsulfonyl(C₁-C₄)alkyl. In another alternative, R¹ is —H,(C₁-C₇)alkyl, or —O—(C₁-C₄)alkyl. In another alternative, R¹ is —H or(C₁-C₇)alkyl. In another alternative, R¹ is —H, methyl or ethyl. In yetanother alternative, R¹ is —OCH₃ or —OC(CH₃)₃.

R² is selected from hydrogen, (C₁-C₇)alkyl,(C₃-C₆)cycloalkyl(C₁-C₄)alkyl, (C₁-C₇)alkoxy(C₁-C₄)alkyl,(C₃-C₆)cycloalkoxy(C₁-C₄)alkyl, (C₃-C₆)cycloalkyl, aryl,aryl(C₁-C₄)alkyl, aryloxy(C₁-C₄)alkyl, arylthio(C₁-C₄)alkyl,arylsulfinyl(C₁-C₄)alkyl, arylsulfonyl(C₁-C₄)alkyl and —O—(C₁-C₇)alkyl.Each alkyl, cycloalkyl, alkoxy, cycloalkoxy, aryl, and aryloxy moiety inthe groups represented by R² can be optionally substituted with one ormore independently selected substituents defined above for StructuralFormula (I). Alternatively, R² is selected from hydrogen, (C₁-C₇)alkyl,(C₃-C₆)cycloalkyl(C₁-C₄)alkyl, (C₁-C₇)alkoxy(C₁-C₄)alkyl,(C₃-C₆)cycloalkoxy(C₁-C₄)alkyl, (C₃-C₆)cycloalkyl, aryl,aryl(C₁-C₄)alkyl, aryloxy(C₁-C₄)alkyl, arylthio(C₁-C₄)alkyl,arylsulfinyl(C₁-C₄)alkyl and arylsulfonyl(C₁-C₄)alkyl. Alternatively, R²is selected from (C₁-C₇)alkyl, (C₃-C₆)cycloalkyl(C₁-C₄)alkyl,(C₁-C₇)alkoxy(C₁-C₄)alkyl, phenyl, phenyl(C₁-C₄)alkyl, (C₃-C₆)cycloalkyland halo(C₁-C₄)alkyl, wherein each alkyl, alkoxy and cycloalkyl moietyin the groups represented by R² is optionally substituted with one ormore substituents independently selected from the group consisting of(C₁-C₄)alkyl and halo; and each phenyl moiety in the groups representedby R² is optionally substituted with one or more substituentsindependently selected from the group consisting of (C₁-C₄)alkyl, halo,(C₁-C₄)alkoxy, (C₁-C₄)alkoxy(C₁-C₄)alkyl, —CN, halo(C₁-C₄)alkyl, andhalo(C₁-C₄)alkoxy. In another alternative, R² is selected from the groupconsisting of cyclopropyl, cyclobutyl, cyclopentyl, cyclopropylmethyl,cyclobutylmethyl, phenyl, benzyl, —(CH₂)₂—O—CH₃, —(CH₂)₃—OCH₃, —C(CH₃)₃,—CH(CH₃)₂, —CH₂C(CH₃)₃, —CH₂CH(CH₃)₂, —CH₂—CF₃, —(CH₂)₂—CH₂F, and—(CH₂)_(n)CH₃, wherein n is 0, 1, 2, 3, 4, 5 or 6; and wherein thephenyl or benzyl group represented by R² is optionally substituted withone or more substituents independently selected from the groupconsisting of (C₁-C₄)alkyl, halo, (C₁-C₄)alkoxy,(C₁-C₄)alkoxy(C₁-C₄)alkyl, —CN, halo(C₁-C₄)alkyl, and halo(C₁-C₄)alkoxy.In another alternative, R² is a phenyl or benzyl group optionallysubstituted with one or more substituents independently selected fromthe group consisting of (C₁-C₄)alkyl, halo, (C₁-C₄)alkoxy,(C₁-C₄)alkoxy(C₁-C₄)alkyl, —CN, halo(C₁-C₄)alkyl, and halo(C₁-C₄)alkoxy.In another alternative, R² is unsubstituted phenyl or benzyl. In anotheralternative, R² is selected from cyclopropyl, cyclopropylmethyl,cyclobutyl, cyclopentyl,

cyclohexyl, —(CH₂)₂—O—CH₃, —C(CH₃)₃, —CH(CH₃)₂, —CH₂—CF₃, —CH₂CH(CH₃)₂,—CH₃ and —CH₂CH₃.

Alternatively, R¹ and R² taken together with the nitrogen atom to whichthey are bonded can also form a monocyclic or bicyclic heteroaryl, or amonocyclic, fused bicyclic, bridged bicyclic or spiro bicyclicheterocycle, wherein the heteroaryl or heterocycle optionally containsone or two additional heteroatoms independently selected from N, O and Sin addition to the N atom to which R₁ and R₂ are bonded. The heteroarylor heterocycle can be optionally substituted with one or moreindependently selected substituents described above for StructuralFormula (I). Alternatively, the heteroaryl or heterocycle contains oneadditional heteroatom selected from N, O and S. Alternatively, R¹ and R²taken together with the nitrogen atom to which they are bonded form aheterocycle selected from the group consisting of azetidine,pyrrolidine, morpholine, piperidine, octahydrocyclopenta[c]pyrrol,isoindoline, and azabicyclo[3.1.0]hexane, wherein the heterocycle isoptionally substituted with one or more substituents independentlyselected from the group consisting of (C₁-C₄)alkyl, halogen, —OH,(C₁-C₄)alkoxy, (C₁-C₄)alkylthio, (C₁-C₄)alkylsulfinyl,(C₁-C₄)alkylsulfonyl, (C₁-C₄)alkoxy(C₁-C₄)alkyl, and —N(R³)(R⁴). In amore specific embodiment, these heterocycles are optionally substitutedwith one or more substituents independently selected from the groupconsisting of halogen, (C₁-C₄)alkoxy, hydroxy, (C₁-C₄)alkoxy(C₁-C₄)alkyland —N(R³)(R⁴). In another alternative embodiment, these heterocyclesare optionally substituted with one or more substituents independentlyselected from the group consisting of halogen, methoxy, hydroxy,methoxymethyl and dimethylamino. Alternatively, R¹ and R² taken togetherwith the nitrogen atom to which they are bonded form a ring selectedfrom pyrrolidinyl, morpholinyl, azetidinyl, piperidinyl,octahydrocyclopenta[c]pyrrolyl, isoindolinyl, indazolyl, imidazolyl,pyrazolyl, triazolyl, and tetrazolyl, wherein the ring formed by R¹ andR² taken together with the nitrogen atom to which they are bonded isoptionally substituted with halogen, (C₁-C₄)alkoxy, hydroxy,(C₁-C₄)alkoxy(C₁-C₄)alkyl and —N(R³)(R⁴). More specifically, the ringformed by R¹ and R² taken together with the nitrogen atom to which theyare bonded is optionally substituted with fluoro, —OH, —OCH₃, orN(CH₃)₂.

R³ and R⁴ are each independently selected from the group consisting of—H and (C₁-C₄)alkyl, wherein the (C₁-C₄)alkyl represented by R³ and R⁴is optionally substituted with one or more substituents independentlyselected from the group consisting of (C₁-C₄)alkyl, halo, —OH,(C₁-C₄)alkoxy, and (C₁-C₄)alkoxy(C₁-C₄)alkyl. Alternatively, R³ and R⁴are both methyl. In another alternative, R³ and R⁴ are both —H. In yetanother alternative, R³ and R⁴ are each unsubstituted (C₁-C₄)alkyl.

In a first alternative embodiment, the compound of the present inventionis represented by Structural Formula (I) or (II), or a pharmaceuticallyacceptable salt thereof, wherein:

R¹ is —H or (C₁-C₇)alkyl; and

R² is selected from (C₁-C₇)alkyl, (C₃-C₆)cycloalkyl(C₁-C₄)alkyl,(C₁-C₇)alkoxy(C₁-C₄)alkyl, phenyl, phenyl(C₁-C₄)alkyl, (C₃-C₆)cycloalkyland halo(C₁-C₄)alkyl, wherein each alkyl, alkoxy or cycloalkyl moiety inthe groups represented by R² is optionally substituted with one or moresubstituents independently selected from the group consisting of(C₁-C₄)alkyl and halo; and each phenyl moiety in the groups representedby R² is optionally substituted with one or more substituentsindependently selected from the group consisting of (C₁-C₄)alkyl, halo,(C₁-C₄)alkoxy, (C₁-C₄)alkoxy(C₁-C₄)alkyl, —CN, halo(C₁-C₄)alkyl, andhalo(C₁-C₄)alkoxy. Alternatively, R² is selected from the groupconsisting of cyclopropyl, cyclobutyl, cyclopentyl, cyclopropylmethyl,cyclobutylmethyl, phenyl,

benzyl, —(CH₂)₂—O—CH₃, —(CH₂)₃—OCH₃, —C(CH₃)₃, —CH(CH₃)₂, —CH₂C(CH₃)₃,—CH₂CH(CH₃)₂, —CH₂—CF₃, —(CH₂)₂—CH₂F, and —(CH₂)_(n)CH₃, wherein n is 0,1, 2, 3, 4, 5 or 6; and wherein the phenyl or benzyl group representedby R² is optionally substituted with one or more substituentsindependently selected from the group consisting of (C₁-C₄)alkyl, halo,(C₁-C₄)alkoxy, (C₁-C₄)alkoxy(C₁-C₄)alkyl, —CN, halo(C₁-C₄)alkyl, andhalo(C₁-C₄)alkoxy. In another alternative, the phenyl or benzyl grouprepresented by R² is unsubstituted. In yet another alternative, R² isselected from cyclopropyl, cyclopropylmethyl, cyclobutyl, cyclopentyl,cyclohexyl, —(CH₂)₂—O—CH₃, —C(CH₃)₃, —CH(CH₃)₂, —CH₂—CF₃, —CH₂CH(CH₃)₂,—CH₃ and —CH₂CH₃.

In a second alternative embodiment, for compounds represented byStructural Formula (I) or (II), R¹ is hydrogen, methyl or ethyl; andvalues and alternative values for R² are as described above for thefirst alternative embodiment.

In a third alternative embodiment, for compounds represented byStructural Formula (I) or (II), R¹ is hydrogen, (C₁-C₄)alkyl or—O—(C₁-C₄)alkyl; R² is selected from (C₁-C₇)alkyl,(C₃-C₆)cycloalkyl(C₁-C₄)alkyl, (C₁-C₇)alkoxy(C₁-C₄)alkyl, phenyl,(C₃-C₆)cycloalkyl, and fluoro(C₁-C₄)alkyl; or R¹ and R² taken togetherwith the nitrogen atom to which they are bonded form a ring selectedfrom pyrrolidinyl, morpholinyl, azetidinyl, piperidinyl,octahydrocyclopenta[c]pyrrolyl, isoindolinyl, indazolyl, imidazolyl,pyrazolyl, triazolyl, and tetrazolyl, wherein the ring formed by R¹ andR² taken together with the nitrogen atom to which they are bonded isoptionally substituted with fluoro, —OH, —OCH₃, or N(CH₃)₂. Morespecifically, R¹ is hydrogen, methyl, ethyl, methoxy or tert-butoxy.

In a fourth alternative embodiment, for compounds represented byStructural Formula (I) or (II), R¹ hydrogen, methyl, or ethyl; R² isselected from methyl, ethyl, n-propyl, isopropyl, n-butyl,2,2-dimethylpropyl, t-butyl, isobutyl, n-pentyl, (C₄-C₆)cycloalkyl,(C₃-C₅)cycloalkylmethyl, methoxyethyl, and 2-fluoroethyl; or R¹ and R²taken together with the nitrogen atom to which they are bonded form aring selected from azetidinyl, pyrrolidinyl, piperidinyl, tetrazolyl, oroctahydrocyclopenta[c]pyrrolyl, and wherein the ring formed by R¹ and R²taken together with the nitrogen atom to which they are bonded isoptionally substituted with fluoro.

In a fifth alternative embodiment, for compounds represented byStructural Formula (A), when X is hydrogen, Y is selected from hydrogen,—(C₁-C₄)alkyl, —(C₁-C₄)alkylene-N(R^(A))(R^(B)),—(C₁-C₄)alkylene-N(R^(F))—C(O)—[C(R^(D))(R^(E))]₀₋₄—N(R^(A))(R^(B)),—CH═N—OR^(A), —N(R^(A))(R^(B)),—N(R^(F′))—C(O)—[C(R^(D))(R^(E))]₁₋₄—N(R^(A))(R^(B)),—NH—C(O)—C(R^(D′))(R^(E))—N(R^(A))(R^(B)),—N(R^(F))—C(O)—N(R^(A))(R^(A)), —N(R^(F))—C(O)—(C₁-C₆)alkyl, —N(R^(F))—C(O)-heterocyclyl, —N(R^(F))—C(O)-heteroaryl, —N(R^(F))—C(O)-carbocyclyl,—N(R^(F))—C(O)-aryl —N(R^(F))—S(O)_(m)—(C₁-C₄)alkylene-N(R^(A))(R^(B)),—N(R^(F))—S(O)_(m)—(C₁-C₄)alkylene-carbocyclyl,—N(R^(F))—S(O)_(m)—(C₁-C₄)alkylene-aryl;

R^(D′) is selected from (C₁-C₆)alkyl, carbocyclyl, aryl, heteroaryl,heterocyclyl, and a naturally occurring amino acid side chain moiety, or

R^(D′) and R^(E) taken together with the carbon atom to which they arebound form a 3-7 membered carbocyclyl, or a 4-7 membered heterocyclyl,wherein the heterocyclyl formed by R^(D′) and R^(E) optionally comprisesone to two additional heteroatoms independently selected from N, S andO; and

R^(F′) is selected from (C₁-C₇)alkyl, carbocyclyl, aryl and heteroaryl.Values and alternative values for the remainder of the variables are asdescribed above for Structural Formula (A). Alternatively, R^(F′) isselected from (C₁-C₄)alkyl and phenyl and the remainder of the variablesare as described above in the fifth alternative embodiment.

In a sixth alternative embodiment, for compounds represented byStructural Formula (A), X is selected from hydrogen, methyl, ethyl andphenyl; and Y is selected from hydrogen, —(C₁-C₄

alkyl), —(C₁-C₄)alkylene-N(R^(A))(R^(B)),—(C₁-C₄)alkylene-NH—C(O)—[CH₂]₀₋₁—N(R^(A))(R^(B)), —N (R^(A))(R^(B)),—NH—C(O)-carbocyclyl, —NH—C(O)-aryl, —NH—C(O)-heterocyclyl,—NH—C(O)-heteroaryl, —NH—C(O)—N(R^(A))(R^(A)),—N(R^(F′))—C(O)—CH₂—N(R^(A))(R^(B)),—NH—C(O)—C(R^(D′))(R^(E))—N(R^(A))(R^(B)) and—NH—S(O)_(m)—(C₁-C₄)alkylene-N(R^(A))(R^(B)); or

X is selected from methyl, ethyl and phenyl; and Y

is —NH—C(O)—CH₂—N(R^(A))(R^(B)), wherein:

each R^(A) is independently selected from hydrogen and methyl;

R^(B) is selected from hydrogen,

(C₁-C₇)alkyl, —(C₀-C₆)alkylene-carbocyclyl, —(C₀-C₆)alkylene-aryl,—(C₀-C₆)alkylene-heteroaryl, —S(O)_(m)—(C₁-C₆)alkyl,—(C₀-C₄)alkylene-S(O)_(m)-carbocyclyl, —(C₀-C₄)alkylene-S(O)_(m)-aryl,—(C₀-C₄)alkylene-S(O)_(m)-heterocycle and—(C₀-C₄)alkylene-S(O)_(m)-heteroaryl; or

R^(A) and R^(B) when bound to a common nitrogen atom are taken togetherwith the nitrogen atom to form a heterocycle, wherein the heterocycle isoptionally substituted with ═O and —N(R^(G))(R^(G));

R^(D′) and R^(E) are taken together with the carbon atom to which theyare bound form a (C₃-C₇)cycloalkyl; and

m is 1 or 2;

each carbocyclyl, aryl, heterocyclyl or heteroaryl is optionally andindependently substituted with one or more substituents independentlyselected from halo, —(C₁-C₄)alkyl, halo-substituted —(C₁-C₄)alkyl,—O—(C₁-C₄)alkyl, and —N(R^(G))(R^(G));

each alkyl portion in the group represented by Y or R^(B) is optionallyand independently substituted with one or more substituentsindependently selected from halo, —OH, and —N(R^(G))(R^(G)), wherein

R^(G) is hydrogen or (C₁-C₄)alkyl, and wherein each alkyl in the grouprepresented by R^(G) is optionally and independently substituted with(C₃-C₆) cycloalkyl. The remainder of the variables are as describedabove in the fifth alternative embodiment.

In a seventh alternative embodiment, for compounds represented byStructural Formula (A), X is selected from hydrogen and methyl; and

Y is selected

from —N(R^(A))(R^(B)), —N(H)—C(O)-carbocyclyl, —N(H)—C(O)-aryl,—N(H)—C(O)-heterocycle, and —N(H)—C(O)-heteroaryl; or

X is methyl; and

Y is —NH—C(O)—CH₂—N(R^(A))(R^(B)), wherein:

R^(A) is hydrogen; and

R^(B) is selected from (C₁-C₄)alkyl, and —S(O)₂—CH₃; or R^(A) and R^(B)taken together to form 4-7 membered heterocyclic ring; wherein eachcarbocyclyl, aryl, heteroaryl, or heterocycle is optionally andindependently substituted with one or more substituents independentlyselected from —CH₃, fluoro, and —N(CH₃)₂.

In a eighth alternative embodiment, for compounds represented byStructural Formula (A), Y is

wherein ring A represents a 4-7 membered heterocyclyl; and R^(3′) ishydrogen or (C₁-C₆)alkyl. Values and alternative values for theremainder of the variables are as described above for Structural Formula(A). More specifically, ring A is selected from the group consisting ofazetidinyl, pyrrolidinyl, piperidinyl, oroctahydrocyclopenta[c]pyrrolyl, each of which is optionally substitutedwith one or more substitutents independently selected from the groupconsisting of halo, —(C₁-C₄)alkyl, halo-substituted-(C₁-C₄)alkyl (e.g.,—CF₃), —OH, —O—(C₁-C₄)alkyl, or —N(R^(G))(R^(G)), wherein R^(G) ishydrogen or (C₁-C₄)alkyl. Even more specifically, ring A described aboveis optionally substituted with one or more fluoro.

In a ninth alternative embodiment, for compounds represented byStructural Formula (A), Y is —NH—C(O)-heteroaryl. Values and alternativevalues for the remainder of the variables are as described above forStructural Formula (A). More specifically, the heteroaryl in—NH—C(O)-heteroaryl is selected from the group consisting of thienyl,pyridinyl, pyrrolyl, oxazolyl, pyrazolyl and thiazolyl, each of which isoptionally substituted with one or more substituents independentlyselected from —(C₁-C₄)alkyl, halo-substituted-(C₁-C₄)alkyl (e.g., —CF₃),—OH, —O—(C₁-C₄)alkyl, and —N(R^(G))(R^(G)), wherein R^(G) is hydrogen or(C₁-C₄)alkyl. More specifically, the pyrrolyl and pyrazolyl areoptionally substituted with a methyl group on the N atom in the ring.

In a tenth alternative embodiment, for compounds represented byStructural Formula (A), Y is —NH—C(O)-phenyl. Values and alternativevalues for the remainder of the variables are as described above forStructural Formula (A). More specifically, the phenyl in —NH—C(O)-phenylis optionally substituted with one or more substitutents independentlyselected from —(C₁-C₄)alkyl, halo-substituted-(C₁-C₄)alkyl (e.g., —CF₃),—OH, —O—(C₁-C₄)alkyl, and —N(R^(G))(R^(G)), wherein R^(G) is hydrogen or(C₁-C₄)alkyl. More specifically, the phenyl in —NH—C(O)-phenyl isoptionally substituted with one or more substitutents independentlyselected from —CF₃, —OCH₃ and —N(CH₃)₂.

In a eleventh alternative embodiment, for compounds represented byStructural Formula (A), Y is represented by —NH—S(O)₂—(C₁-C₆)alkyl,—NH—S(O)₂-phenyl, —NH—S(O)₂-heteroaryl. Values and alternative valuesfor the remainder of the variables are as described above for StructuralFormula (A). More specifically, the phenyl, heteroaryl or alkyl in thegroup represented by Y is optionally substituted with one or moresubstituents independently selected from —(C₁-C₄)alkyl,halo-substituted-(C₁-C₄)alkyl (e.g., —CF₃), —OH, —O—(C₁-C₄)alkyl, and—N(R^(G))(R^(G)), wherein R^(G) is hydrogen or (C₁-C₄)alkyl.

In a twelfth alternative embodiment, for compounds represented byStructural Formula (A), Y is represented by —N(R^(A))(R^(B)), whereinR^(A) and R^(B) are each independently selected from hydrogen,(C₁-C₇)alkyl, —(C₁-C₄)alkyl-(C₃-C₆)cycloalkyl, wherein the (C₁-C₇)alkylis optionally substituted with —N(R^(G))(R^(G)), wherein R^(G) ishydrogen or (C₁-C₄)alkyl. Values and alternative values for theremainder of the variables are as described above for Structural Formula(A).

In a thirteenth alternative embodiment, for compounds represented byStructural Formula (A), Y is represented by —CH₂—N(R^(A))(R^(B)),wherein R^(A) and R^(B) are each independently selected from hydrogen,(C₁-C₇)alkyl, —(C₁-C₄)alkyl-(C₃-C₆)cycloalkyl, or R^(A) and R^(B) takentogether with the nitrogen atom to which they are bound form aheterocyclyl, wherein the (C₁-C₇)alkyl represented by R^(A) or R^(B) isoptionally and independently substituted with —N(R^(G))(R^(G)), whereinR^(G) is hydrogen or (C₁-C₄)alkyl, and the (C₁-C₄)alkyl represented byR^(G) is optionally substituted with —F. Values and alternative valuesfor the remainder of the variables are as described above for StructuralFormula (A).

In a fourteenth alternative embodiment, for compounds represented byStructural Formula (A), Y is represented by—CH₂—NH—C(O)—(CH₂)₀₋₁—N(R^(A))(R^(B)), wherein R^(A) and R^(B) are eachindependently selected from hydrogen, (C₁-C₇)alkyl,—(C₁-C₄)alkyl-(C₃-C₆)cycloalkyl. Values and alternative values for theremainder of the variables are as described above for Structural Formula(A).

The compound of the present invention is exemplified by the compoundsshown in the Table below or a pharmaceutically acceptable salt thereofand compounds described in Examples 1-12 below or a pharmaceuticallyacceptable salt thereof:

Compound No. Chemical Structure 11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

“Alkyl” means a saturated aliphatic branched or straight-chainmonovalent hydrocarbon radical having the specified number of carbonatoms. Thus, “(C₁-C₇)alkyl” means a radical having from 1-7 carbon atomsin a linear or branched arrangement. “(C₁-C₇)alkyl” includes methyl,ethyl, propyl, butyl, pentyl, hexyl and heptyl. Suitable substitutionsfor a “substituted alkyl” include, but are not limited to, -halogen,—OH, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)alkylthio,(C₁-C₄)alkylsulfinyl, (C₁-C₄)alkylsulfonyl, (C₁-C₄)alkoxy(C₁-C₄)alkyl,and —N(R³)(R⁴), wherein R³ and R⁴ are as described above.

“Cycloalkyl” means a saturated aliphatic cyclic hydrocarbon radicalhaving the specified number of carbon atoms. (C₃-C₆)cycloalkyl includescyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Suitablesubstituents for a “substituted cycloalkyl” include halogen, —OH,(C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)alkylthio, (C₁-C₄)alkylsulfinyl,(C₁-C₄)alkylsulfonyl, (C₁-C₄)alkoxy(C₁-C₄)alkyl, and —N(R³)(R⁴), whereinR³ and R⁴ are as described above.

“Heterocycle” means a 4-12 membered partially unsaturated or saturatedheterocyclic ring containing 1, 2, or 3 heteroatoms independentlyselected from N, O or S. When one heteroatom is S, it can be optionallymono- or di-oxygenated (i.e. —S(O)— or —S(O)₂—). The heterocycle can bemonocyclic, fused bicyclic, bridged bicyclic, or spiro bicyclic.

Examples of monocyclic heterocycle include, but not limited to,azetidine, pyrrolidine, piperidine, piperazine, hexahydropyrimidine,tetrahydrofuran, tetrahydropyran, morpholine, thiomorpholine,thiomorpholine 1,1-dioxide, tetrahydro-2H-1,2-thiazine,tetrahydro-2H-1,2-thiazine 1,1-dioxide, isothiazolidine, isothiazolidine1,1-dioxide.

A fused bicyclic heterocycle has two rings which have two adjacent ringatoms in common. The first ring is a monocyclic heterocycle and thesecond ring is a cycloalkyl, partially unsaturated carbocycle, phenyl,heteroaryl or a monocyclic heterocycle. For example, the second ring isa (C₃-C₆)cycloalkyl, such as cyclopropyl, cyclobutyl, cyclopentyl andcyclohexyl. Alternatively, the second ring is phenyl. Example of fusedbicyclic heterocycles includes, but not limited to, indoline,isoindoline, 2,3-dihydro-1H-benzo[d]imidazole,2,3-dihydrobenzo[d]oxazole, 2,3-dihydrobenzo[d]thiazole,octahydrobenzo[d]oxazole, octahydro-1H-benzo[d]imidazole,octahydrobenzo[d]thiazole, octahydrocyclopenta[c]pyrrole,3-azabicyclo[3.1.0]hexane, and 3-azabicyclo[3.2.0]heptane.

A spiro bicyclic heterocycle has two rings which have only one ring atomin common. The first ring is a monocyclic heterocycle and the secondring is a cycloalkyl, partially unsaturated carbocycle or a monocyclicheterocycle. For example, the second ring is a (C₃-C₆)cycloalkyl.Example of spiro bicyclic heterocycle includes, but not limited to,azaspiro[4.4]nonane, 7-azaspiro[4.4]nonane, azasprio[4.5]decane,8-azaspiro[4.5]decane, azaspiro[5.5]undecane, 3-azaspiro[5.5]undecaneand 3,9-diazaspiro[5.5]undecane.

A bridged bicyclic heterocycle has two rings which have three or moreadjacent ring atoms in common. The first ring is a monocyclicheterocycle and the other ring is a cycloalkyl (such as(C₃-C₆)cycloalkyl), partially unsaturated carbocycle or a monocyclicheterocycle. Examples of bridged bicyclic heterocycles include, but arenot limited to, azabicyclo[3.3.1]nonane, 3-azabicyclo[3.3.1]nonane,azabicyclo[3.2.1]octane, 3-azabicyclo[3.2.1]octane,6-azabicyclo[3.2.1]octane and azabicyclo[2.2.2]octane,2-azabicyclo[2.2.2]octane.

When the heterocycle contains a N atom other than the nitrogen atom towhich R¹ and R² are bonded, the N atom can be substituted with H, alkyl,cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, each of which can be optionally substituted withhalogen, hydroxy, alkoxy, haloalkyl, alkyl, etc. The heterocycle can beoptionally substituted with an oxo group (C═O) and oxo substitutedheterocyclic rings include, but are not limited to, thiomorpholine1-oxide, thiomorpholine 1,1-dioxide, tetrahydro-2H-1,2-thiazine1,1-dioxide, and isothiazolidine 1,1-dioxide, pyrrolidin-2-one,piperidin-2-one, piperazin-2-one, and morpholin-2-one. Other optionalsubstituents for a heterocycle include (C₁-C₄)alkyl, halo, —OH,(C₁-C₄)alkoxy, (C₁-C₄)alkylthio, (C₁-C₄)alkylsulfinyl,(C₁-C₄)alkylsulfonyl, (C₁-C₄)alkoxy(C₁-C₄)alkyl, —N(R³)(R⁴), —CN,halo(C₁-C₄)alkyl, and halo(C₁-C₄)alkoxy.

“Heteroaryl” means a 5-12 membered monovalent heteroaromatic monocyclicor bicylic ring radical. A herteroaryl contains 1, 2 or 3 heteroatomsindependently selected from N, O, and S. Heteroaryls include, but arenot limited to pyrrole, imidazole, pyrazole, oxazole, isoxazole,thiazole, isothiazole, 1,2,3-triazole, 1,2,4-triazole, 1,3,4-oxadiazole,1,2,5-thiadiazole, 1,2,5-thiadiazole 1-oxide, 1,2,5-thiadiazole1,1-dioxide, 1,3,4-thiadiazole, pyridine, pyrazine, pyrimidine,pyridazine, 1,2,4-triazine, 1,3,5-triazine, and tetrazole. Bicyclicheteroaryl rings include, but are not limited to, bicyclo[4.4.0] andbicyclo[4.3.0] fused ring systems such as indolizine, indole, isoindole,indazole, benzimidazole, benzthiazole, purine, quinoline, isoquinoline,cinnoline, phthalazine, quinazoline, quinoxaline, 1,8-naphthyridine, andpteridine.

“Carbocycle” means 4-12 membered saturated or unsaturated aliphaticcyclic hydrocarbon ring.

“Alkoxy” means an alkyl radical attached through an oxygen linking atom.“Alkoxy” can also be depicted as —O-alkyl. For example, (C₁-C₄)-alkoxycan also depicted as —O—(C₁-C₄)alkyl. “(C₁-C₄)-alkoxy” includes methoxy,ethoxy, propoxy, and butoxy.

“Alkylthio” means an alkyl radical attached through a sulfur linkingatom. “Alkylthio” can also be depicted as —S-alkyl. For example,“(C₁-C₄)alkylthio” can be depicted as —S—(C₁-C₄)alkyl.“(C₁-C₄)alkylthio” include methylthio, ethylthio, propylthio andbutylthio.

“Alkylsulfinyl” means an alkyl radical attached through a —S(O)— linkinggroup. “Alkylsulfinyl” can be depicted as —S(O)-alkyl. For example,“(C₁-C₄)alkylsulfinyl” can be depicted as —S(O)—(C₁-C₄)alkyl.“(C₁-C₄)alkylsulfinyl” include methylsulfinyl, ethylsulfinyl,propylsulfinyl and butylsulfinyl.

“Alkylsulfonyl” means an alkyl radical attached through a —S(O)₂—linking group. “Alkylsulfonyl” can be depicted as —S(O)₂-alkyl. Forexample, “(C₁-C₄)alkylsulfinyl” can be depicted as —S(O)₂—(C₁-C₄)alkyl.“(C₁-C₄)alkylsulfonyl” include methylsulfonyl, ethylsulfonyl,propylsulfonyl and butylsulfonyl.

Haloalkyl and halocycloalkyl include mono, poly, and perhaloalkyl groupswhere each halogen is independently selected from fluorine, chlorine,and bromine. Haloalkyl and halocycloalkyl can also be referred ashalo-substituted alkyl and halo-substituted cycloalkyl, respectively.

“Cycloalkoxy” means a cycloalkyl radical attached through an oxygenlinking atom. “Cycloalkoxy” can also be depicted as —O-cycloalkyl. Forexample, “(C₃-C₆)cycloalkoxy” can be depicted as —O—(C₃-C₆)cycloalkyl.“(C₃-C₆)cycloalkoxy” includes cyclopropyloxy, cyclobutyloxy,cyclopentyloxy and cyclohexyloxy.

“Aryl” means an aromatic monocyclic or polycyclic (e.g. bicyclic ortricyclic) carbocyclic ring system. In one embodiment, “aryl” is a 6-12membered monocylic or bicyclic systems. Aryl systems include, but notlimited to, phenyl, naphthalenyl, fluorenyl, indenyl, azulenyl, andanthracenyl.

“Aryloxy” means an aryl moiety attached through an oxygen linking atom.“Aryloxy” can be also depicted as —O-aryl. Aryloxy includes, but notlimited to, phenoxy.

“Arylthio” means an aryl moiety attached through a sulfur linking atom.“Arylthio” can be also depicted as —S-aryl. Arylthio includes, but notlimited to, phenylthio.

“Arylsulfinyl” means an aryl moiety attached through a —S(O)— linkinggroup. “Arylsulfinyl” can be also depicted as —S(O)-aryl. Arylsulfinylincludes, but not limited to, phenylsulfinyl.

“Arylsulfonyl” means an aryl moiety attached through a —S(O)₂— linkinggroup. “Arylsulfonyl”” can be also depicted as —S(O)₂-aryl. Arylsulfonylincludes, but not limited to, phenylsulfonyl.

“Hetero” refers to the replacement of at least one carbon atom member ina ring system with at least one heteroatom selected from N, S, and O.“Hetero” also refers to the replacement of at least one carbon atommember in a acyclic system. A hetero ring system or a hetero acyclicsystem may have 1, 2, or 3 carbon atom members replaced by a heteroatom.

“Halogen” or “halo” used herein refers to fluorine, chlorine, bromine,or iodine.

As used herein, cycloalkylalkyl can be depicted as -alkylene-cycloalkyl.For example, (C₃-C₆)cycloalkyl(C₁-C₄)alkyl can be depicted as—(C₁-C₄)alkylene-(C₃-C₆)cycloalkyl.

As use herein, alkoxyalkyl can be depicted as -alkylene-O-alkyl. Forexample, (C₁-C₇)alkoxy(C₁-C₄)alkyl can be depicted as—(C₁-C₄)alkylene-O—(C₁-C₇)alkyl.

As use herein, cycloalkoxyalkyl can be depicted as-alkylene-O-cycloalkyl. For example, (C₃-C₆)cycloalkoxy(C₁-C₄)alkyl canbe depicted as —(C₁-C₄)alkylene-O—(C₃-C₆)alkyl.

As use herein, arylalkyl can be depicted as -alkylene-aryl. For example,aryl(C₁-C₄)alkyl can be depicted as —(C₁-C₄)alkylene-aryl.

As used herein, aryloxyalkyl can be depicted as -alkylene-O-aryl. Forexample, aryloxy(C₁-C₄)alkyl can be depicted as —(C₁-C₄)alkylene-O-aryl.

As used herein, arylthioalkyl can be depicted as -alkylene-S-aryl. Forexample, arylthio(C₁-C₄)alkyl can be depicted as—(C₁-C₄)alkylene-S-aryl.

As used herein, arylsufinylalkyl can be depicted as -alkylene-S(O)-aryl.For example, arylsufinyl(C₁-C₄)alkyl can be depicted as—(C₁-C₄)alkylene-S(O)-aryl.

As used herein, arylsulfonylalkyl can be depicted as-alkylene-S(O)₂-aryl. For example, arylsulfonyl(C₁-C₄)alkyl can bedepicted as —(C₁-C₄)alkylene-S(O)₂-aryl.

Another embodiment of the present invention is a pharmaceuticalcomposition comprising one or more pharmaceutically acceptable carrierand/or diluent and a compound disclosed herein or a pharmaceuticallyacceptable salt thereof.

“Pharmaceutically acceptable carrier” and “pharmaceutically acceptablediluent” means non-therapeutic components that are of sufficient purityand quality for use in the formulation of a composition of the inventionthat, when appropriately administered to an animal or human, typicallydo not produce an adverse reaction, and that are used as a vehicle for adrug substance (i.e. a compound of the present invention).

Pharmaceutically acceptable salts of the compounds of the presentinvention are also included. For example, an acid salt of a compound ofthe present invention containing an amine or other basic group can beobtained by reacting the compound with a suitable organic or inorganicacid, resulting in pharmaceutically acceptable anionic salt forms.Examples of anionic salts include the acetate, benzenesulfonate,benzoate, bicarbonate, bitartrate, bromide, calcium edetate, camsylate,carbonate, chloride, citrate, dihydrochloride, edetate, edisylate,estolate, esylate, fumarate, glyceptate, gluconate, glutamate,glycollylarsanilate, hexylresorcinate, hydrobromide, hydrochloride,hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate,maleate, mandelate, mesylate, methylsulfate, mucate, napsylate, nitrate,pamoate, pantothenate, phosphate/diphosphate, polygalacturonate,salicylate, stearate, subacetate, succinate, sulfate, tannate, tartrate,teoclate, tosylate, and triethiodide salts.

Salts of the compounds of the present invention containing a carboxylicacid or other acidic functional group can be prepared by reacting with asuitable base. Such a pharmaceutically acceptable salt may be made witha base which affords a pharmaceutically acceptable cation, whichincludes alkali metal salts (especially sodium and potassium), alkalineearth metal salts (especially calcium and magnesium), aluminum salts andammonium salts, as well as salts made from physiologically acceptableorganic bases such as trimethylamine, triethylamine, morpholine,pyridine, piperidine, picoline, dicyclohexylamine,N,N′-dibenzylethylenediamine, 2-hydroxyethylamine,bis-(2-hydroxyethyl)amine, tri-(2-hydroxyethyl)amine, procaine,dibenzylpiperidine, dehydroabietylamine, N,N′-bisdehydroabietylamine,glucamine, N-methylglucamine, collidine, quinine, quinoline, and basicamino acids such as lysine and arginine.

The invention also includes various isomers and mixtures thereof.Certain of the compounds of the present invention may exist in variousstereoisomeric forms. Stereoisomers are compounds which differ only intheir spatial arrangement. Enantiomers are pairs of stereoisomers whosemirror images are not superimposable, most commonly because they containan asymmetrically substituted carbon atom that acts as a chiral center.“Enantiomer” means one of a pair of molecules that are mirror images ofeach other and are not superimposable. Diastereomers are stereoisomersthat are not related as mirror images, most commonly because theycontain two or more asymmetrically substituted carbon atoms. “R” and “S”represent the configuration of substituents around one or more chiralcarbon atoms. When a chiral center is not defined as R or S, either apure enantiomer or a mixture of both configurations is present.

“Racemate” or “racemic mixture” means a compound of equimolar quantitiesof two enantiomers, wherein such mixtures exhibit no optical activity;i.e., they do not rotate the plane of polarized light.

The compounds of the invention may be prepared as individual isomers byeither isomer-specific synthesis or resolved from an isomeric mixture.Conventional resolution techniques include forming the salt of a freebase of each isomer of an isomeric pair using an optically active acid(followed by fractional crystallization and regeneration of the freebase), forming the salt of the acid form of each isomer of an isomericpair using an optically active amine (followed by fractionalcrystallization and regeneration of the free acid), forming an ester oramide of each of the isomers of an isomeric pair using an optically pureacid, amine or alcohol (followed by chromatographic separation andremoval of the chiral auxiliary), or resolving an isomeric mixture ofeither a starting material or a final product using various well knownchromatographic methods.

When the stereochemistry of a disclosed compound is named or depicted bystructure, the named or depicted stereoisomer is at least 60%, 70%, 80%,90%, 99% or 99.9% by weight pure relative to the other stereoisomers.When a single enantiomer is named or depicted by structure, the depictedor named enantiomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% byweight optically pure. Percent optical purity by weight is the ratio ofthe weight of the enantiomer that is present divided by the combinedweight of the enantiomer that is present and the weight of its opticalisomer.

The present invention also provides a method of treating or preventing asubject with a tetracycline-responsive disease or disorder comprisingadministering to the subject an effective amount of a compound of thepresent invention or a pharmaceutically acceptable salt thereof.

“Tetracycline-responsive disease or disorder” refers to a disease ordisorder that can be treated, prevented, or otherwise ameliorated by theadministration of a tetracycline compound of the present invention.Tetracycline-responsive disease or disorder includes infections, cancer,inflammatory disorders, autoimmune disease, arteriosclerosis, cornealulceration, emphysema, arthritis, osteoporosis, osteoarthritis, multiplesclerosis, osteosarcoma, osteomyelitis, bronchiectasis, chronicpulmonary obstructive disease, skin and eye diseases, periodontitis,osteoporosis, rheumatoid arthritis, ulcerative colitis, prostatitis,tumor growth and invasion, metastasis, diabetes, diabetic proteinuria,panbronchiolitis; aortic or vascular aneurysms, skin tissue wounds, dryeye, bone, cartilage degradation, malaria, senescence, diabetes,vascular stroke, neurodegenerative disorders, cardiac disease, juvenilediabetes, acute and chronic bronchitis, sinusitis, and respiratoryinfections, including the common cold; Wegener's granulomatosis;neutrophilic dermatoses and other inflammatory diseases such asdermatitis herpetiformis, leukocytoclastic vasculitis, bullous lupuserythematosus, pustular psoriasis, erythema elevatum diutinum; vitiligo;discoid lupus erythematosus; pyoderma gangrenosum; pustular psoriasis;blepharitis, or meibomianitis; Alzheimer's disease; degenerativemaculopathy; acute and chronic gastroenteritis and colitis; acute andchronic cystitis and urethritis; acute and chronic dermatitis; acute andchronic conjunctivitis; acute and chronic serositis; uremicpericarditis; acute and chronic cholecystis; cystic fibrosis, acute andchronic vaginitis; acute and chronic uveitis; drug reactions; insectbites; burns and sunburn, bone mass disorder, acute lung injury, chroniclung disorders, ischemia, stroke or ischemic stroke, skin wound, aorticor vascular aneurysm, diabetic retinopathy, hemorrhagic stroke,angiogenesis, and other states for which tetracycline compounds havebeen found to be active (see, for example, U.S. Pat. Nos. 5,789,395;5,834,450; 6,277,061 and 5,532,227, each of which is expresslyincorporated herein by reference).

In addition, a method to treat any disease or disease state that couldbenefit from modulating the expression and/or function of nitric oxide,metalloproteases, proinflammatory mediators and cytokines, reactiveoxygen species, components of the immune response, including chemotaxis,lymphocyte transformation, delayed hypersensitivity, antibodyproduction, phagocytosis, and oxidative metabolism of phagocytes. Amethod to treat any disease or disease state that could benefit frommodulating the expression and/or function of C-reactive protein,signaling pathways (e.g., FAK signaling pathway), and/or augment theexpression of COX-2 and PGE₂ production is covered. A method to treatany disease or disease state that could benefit from inhibition ofneovascularization is covered.

Compounds of the invention can be used to prevent or treat importantmammalian and veterinary diseases such as diarrhea, urinary tractinfections, infections of skin and skin structure, ear, nose and throatinfections, wound infection, mastitis and the like. In addition, methodsfor treating neoplasms using tetracycline compounds of the invention arealso included (van der Bozert et al., Cancer Res., 48: 6686-6690(1988)).

Infections that can be treated using compounds of the invention or apharmaceutically acceptable salt thereof include, but are not limitedto, skin infections, GI infections, urinary tract infections,genito-urinary infections, respiratory tract infections, sinusesinfections, middle ear infections, systemic infections, cholera,influenza, bronchitis, acne, malaria, sexually transmitted diseaseincluding syphilis and gonorrhea, Legionnaires' disease, Lyme disease,Rocky Mountain spotted fever, Q fever, typhus, bubonic plague, gasgangrene, hospital acquired infections, leptospirosis, whooping cough,anthrax and infections caused by the agents responsible forlymphogranuloma venereum, inclusion conjunctivitis, or psittacosis.Infections can be bacterial, fungal, parasitic and viral infections(including those which are resistant to other tetracycline compounds).

In one embodiment, the infection can be caused bacteria. In anotherembodiment, the infection is caused by a Gram-positive bacteria. In aspecific aspect of this embodiment, the infection is caused by aGram-positive bacterium selected from Staphylococcus spp., Streptococcusspp., Propionibacterium spp., Enterococcus spp., Bacillus spp.,Corynebacterium spp., Nocardia spp, Clostridium spp., Actinobacteriaspp., and Listeria spp.

In another embodiment, the infection is caused by a Gram-negativebacterium. In one aspect of this embodiment, the infection is caused bya proteobacteria (e.g., Betaproteobacteria and Gammaproteobacteria),including Escherichia coli, Salmonella, Shigella, otherEnterobacteriaceae, Pseudomonas, Moraxella, Helicobacter,Stenotrophomonas, Bdellovibrio, acetic acid bacteria, Legionella oralpha-proteobacteria such as Wolbachia. In another aspect, the infectionis caused by a Gram-negative bacteria selected from cyanobacteria,spirochaetes, green sulfur or green non-sulfur bacteria. In a specificaspect of this embodiment, the infection is caused by a Gram-negativebacteria selected from Enterobactericeae (e.g., E. coli, Klebsiellapneumonia including those containing extended-spectrum β-lactamasesand/or carbapenemases), Bacteroidaceae (e.g., Bacteroides fagilis).,Vibrionaceae (Vibrio cholerae), Pasteurellae (e.g., Haemophilusinfluenza), Pseudomonadaceae (e.g., Pseudomonas aeruginosa),Neisseriaceae (e.g. Neisseria meningitidis), Rickettsiae, Moraxellaceae(e.g., Moraxella catarrhalis), any species of Proteeae, Acinetobacterspp., Helicobacter spp., and Campylobacter spp.

In a particular embodiment, the infection is caused by Gram-negativebacterium selected from the group consisting of Enterobactericeae (e.g.,E. coli, Klebsiella pneumoniae), Pseudomonas, and Acinetobacter spp.

In another embodiment, the infection is caused by an organism selectedfrom the group consisting of K. pneumoniae, Salmonella, E. hirae, A.baumanii, M. catarrhalis, H. influenzae, P. aeruginosa, E. faecium, E.coli, S. aureus, and E. faecalis.

In another embodiment, the infection is caused by an organism selectedfrom the group consisting of rickettsiae, chlamydiae, Legionella spp.and Mycoplasma spp.

In another embodiment, the infection is caused by an organism resistantto tetracycline or any member of first and second generation oftetracycline antibiotics (e.g., doxycycline or minocycline).

In another embodiment, the infection is caused by an organism resistantto methicillin.

In another embodiment, the infection is caused by an organism resistantto vancomycin.

In another embodiment, the infection is caused by an organism resistantto a quinolone or fluoroquinolone.

In another embodiment, the infection is caused by an organism resistantto tigecycline.

In another embodiment, the infection is caused by a multidrug-resistantpathogen (having intermediate or full resistance to any two or moreantibiotics). In another embodiment the infection is a Bacillusanthracis infection. “Bacillus anthracis infection” includes any state,diseases, or disorders caused or which result from exposure or allegedexposure to Bacillus anthracis or another member of the Bacillus cereusgroup of bacteria. In another embodiment, the infection is caused byBacillus anthracis (anthrax), Yersinia pestis (plague), or Francisellatularensis (tularemia).

In yet another embodiment, the infection can be caused by more than oneorganism described above. Examples of such infections include, but arenot limited to, intra-abdominal infections (often a mixture of agram-negative species like E. coli and an anaerobe like B. fragilis),diabetic foot (various combinations of Streptococcus, Serratia,Staphylococcus and Enterococcus spp., anaerobes (S. E. Dowd, et al.,PloS one 2008; 3:e3326) and respiratory disease (especially in patientsthat have chronic infections like cystic fibrosis—e.g., S. aureus plusP. aeruginosa or H. influenza, atypical pathogens), wounds and abscesses(various gram-negative and gram-positive bacteria, notably MSSA/MRSA,coagulase-negative staphylococci, enterococci, Acinetobacter, Paeruginosa, E. coli, B. fragilis), and bloodstream infections (13% werepolymicrobial (H. Wisplinghoff, et al., Clin. Infect. Dis. 2004;39:311-317)).

In a further embodiment, the tetracycline responsive disease or disorderis not a bacterial infection. In another embodiment, the tetracyclinecompounds of the invention are essentially non-antibacterial. Forexample, non-antibacterial compounds of the invention may have MICvalues greater than about 4 μg/ml (as measured by assays known in theart and/or the assay given in Example 14. In another embodiment, thetetracycline compounds of the invention have both antibacterial andnon-antibacterial effects.

Tetracycline responsive disease or disorder also includes diseases ordisorders associated with inflammatory process associated states (IPAS).The term “inflammatory process associated state” includes states inwhich inflammation or inflammatory factors (e.g., matrixmetalloproteinases (MMPs), nitric oxide (NO), TNF, interleukins, plasmaproteins, cellular defense systems, cytokines, lipid metabolites,proteases, toxic radicals, adhesion molecules, etc.) are involved or arepresent in an area in aberrant amounts, e.g., in amounts which may beadvantageous to alter, e.g., to benefit the subject. The inflammatoryprocess is the response of living tissue to damage. The cause ofinflammation may be due to physical damage, chemical substances,micro-organisms, tissue necrosis, cancer or other agents. Acuteinflammation is short-lasting, lasting only a few days. If it is longerlasting however, then it may be referred to as chronic inflammation.

IPASs include inflammatory disorders. Inflammatory disorders aregenerally characterized by heat, redness, swelling, pain and loss offunction. Examples of causes of inflammatory disorders include, but arenot limited to, microbial infections (e.g., bacterial and fungalinfections), physical agents (e.g., burns, radiation, and trauma),chemical agents (e.g., toxins and caustic substances), tissue necrosisand various types of immunologic reactions.

Examples of inflammatory disorders can be treated using the compounds ofthe invention or a pharmaceutically acceptable salt thereof include, butare not limited to, osteoarthritis, rheumatoid arthritis, acute andchronic infections (bacterial and fungal, including diphtheria andpertussis); acute and chronic bronchitis, sinusitis, and upperrespiratory infections, including the common cold; acute and chronicgastroenteritis and colitis; inflammatory bowel disorder; acute andchronic cystitis and urethritis; vasculitis; sepsis; nephritis;pancreatitis; hepatitis; lupus; inflammatory skin disorders including,for example, eczema, dermatitis, psoriasis, pyoderma gangrenosum, acnerosacea, and acute and chronic dermatitis; acute and chronicconjunctivitis; acute and chronic serositis (pericarditis, peritonitis,synovitis, pleuritis and tendinitis); uremic pericarditis; acute andchronic cholecystis; acute and chronic vaginitis; acute and chronicuveitis; drug reactions; insect bites; burns (thermal, chemical, andelectrical); and sunburn.

IPASs also include matrix metalloproteinase associated states (MMPAS).MMPAS include states characterized by aberrant amounts of MMPs or MMPactivity.

Examples of matrix metalloproteinase associated states (“MMPAS's”) canbe treated using compounds of the invention or a pharmaceuticallyacceptable salt thereof, include, but are not limited to,arteriosclerosis, corneal ulceration, emphysema, osteoarthritis,multiple sclerosis (Liedtke et al., Ann. Neurol. 1998, 44: 35-46;Chandler et al., J. Neuroimmunol. 1997, 72: 155-71), osteosarcoma,osteomyelitis, bronchiectasis, chronic pulmonary obstructive disease,skin and eye diseases, periodontitis, osteoporosis, rheumatoidarthritis, ulcerative colitis, inflammatory disorders, tumor growth andinvasion (Stetler-Stevenson et al., Annu. Rev. Cell Biol. 1993, 9:541-73; Tryggvason et al., Biochim. Biophys. Acta 1987, 907: 191-217; Liet al., Mol. Carcillog. 1998, 22: 84-89)), metastasis, acute lunginjury, stroke, ischemia, diabetes, aortic or vascular aneurysms, skintissue wounds, dry eye, bone and cartilage degradation (Greenwald etal., Bone 1998, 22: 33-38; Ryan et al., Curr. Op. Rheumatol. 1996, 8:238-247). Other MMPAS include those described in U.S. Pat. Nos.5,459,135; 5,321,017; 5,308,839; 5,258,371; 4,935,412; 4,704,383,4,666,897, and RE 34,656, incorporated herein by reference in theirentirety.

In a further embodiment, the IPAS includes disorders described in U.S.Pat. Nos. 5,929,055; and 5,532,227, incorporated herein by reference intheir entirety.

Tetracycline responsive disease or disorder also includes diseases ordisorders associated with NO associated states. The term “NO associatedstates” includes states which involve or are associated with nitricoxide (NO) or inducible nitric oxide synthase (iNOS). NO associatedstate includes states which are characterized by aberrant amounts of NOand/or iNOS. Preferably, the NO associated state can be treated byadministering tetracycline compounds of the invention. The disorders,diseases and states described in U.S. Pat. Nos. 6,231,894; 6,015,804;5,919,774; and 5,789,395 are also included as NO associated states. Theentire contents of each of these patents are hereby incorporated hereinby reference.

Examples of diseases or disorders associated with NO associated statescan be treated using the compounds of the present invention or apharmaceutically acceptable salt thereof include, but are not limitedto, malaria, senescence, diabetes, vascular stroke, neurodegenerativedisorders (Alzheimer's disease and Huntington's disease), cardiacdisease (reperfusion-associated injury following infarction), juvenilediabetes, inflammatory disorders, osteoarthritis, rheumatoid arthritis,acute, recurrent and chronic infections (bacterial, viral and fungal);acute and chronic bronchitis, sinusitis, and respiratory infections,including the common cold; acute and chronic gastroenteritis andcolitis; acute and chronic cystitis and urethritis; acute and chronicdermatitis; acute and chronic conjunctivitis; acute and chronicserositis (pericarditis, peritonitis, synovitis, pleuritis andtendonitis); uremic pericarditis; acute and chronic cholecystis; cysticfibrosis, acute and chronic vaginitis; acute and chronic uveitis; drugreactions; insect bites; burns (thermal, chemical, and electrical); andsunburn.

In another embodiment, the tetracycline responsive disease or disorderis cancer. Examples of cancers that can be treated using the compoundsof the invention or a pharmaceutically acceptable salt thereof includeall solid tumors, i.e., carcinomas e.g., adenocarcinomas, and sarcomas.Adenocarcinomas are carcinomas derived from glandular tissue or in whichthe tumor cells form recognizable glandular structures. Sarcomas broadlyinclude tumors whose cells are embedded in a fibrillar or homogeneoussubstance like embryonic connective tissue. Examples of carcinomas whichmay be treated using the methods of the invention include, but are notlimited to, carcinomas of the prostate, breast, ovary, testis, lung,colon, and breast. The methods of the invention are not limited to thetreatment of these tumor types, but extend to any solid tumor derivedfrom any organ system. Examples of treatable cancers include, but arenot limited to, colon cancer, bladder cancer, breast cancer, melanoma,ovarian carcinoma, prostate carcinoma, lung cancer, and a variety ofother cancers as well. The methods of the invention also cause theinhibition of cancer growth in adenocarcinomas, such as, for example,those of the prostate, breast, kidney, ovary, testes, and colon. In oneembodiment, the cancers treated by methods of the invention includethose described in U.S. Pat. Nos. 6,100,248; 5,843,925; 5,837,696; or5,668,122, incorporated herein by reference in their entirety.

Alternatively, the tetracycline compounds may be useful for preventingor reducing the likelihood of cancer recurrence, for example, to treatresidual cancer following surgical resection or radiation therapy. Thetetracycline compounds useful according to the invention are especiallyadvantageous as they are substantially non-toxic compared to othercancer treatments.

In a further embodiment, the compounds of the invention are administeredin combination with standard cancer therapy, such as, but not limitedto, chemotherapy.

Examples of tetracycline responsive states can be treated using thecompounds of the invention or a pharmaceutically acceptable salt thereofalso include neurological disorders which include both neuropsychiatricand neurodegenerative disorders, but are not limited to, such asAlzheimer's disease, dementias related to Alzheimer's disease (such asPick's disease), Parkinson's and other Lewy diffuse body diseases,senile dementia, Huntington's disease, Gilles de la Tourette's syndrome,multiple sclerosis, amyotrophic lateral sclerosis (ALS), progressivesupranuclear palsy, epilepsy, and Creutzfeldt-Jakob disease; autonomicfunction disorders such as hypertension and sleep disorders, andneuropsychiatric disorders, such as depression, schizophrenia,schizoaffective disorder, Korsakoffs psychosis, mania, anxietydisorders, or phobic disorders; learning or memory disorders, e. g.,amnesia or age-related memory loss, attention deficit disorder,dysthymic disorder, major depressive disorder, mania,obsessive-compulsive disorder, psychoactive substance use disorders,anxiety, phobias, panic disorder, as well as bipolar affective disorder,e. g., severe bipolar affective (mood) disorder (BP-1), bipolaraffective neurological disorders, e. g., migraine and obesity.

Further neurological disorders include, for example, those listed in theAmerican Psychiatric Association's Diagnostic and Statistical manual ofMental Disorders (DSM), the most current version of which isincorporated herein by reference in its entirety.

In another embodiment, the tetracycline responsive disease or disorderis diabetes. Diabetes that can be treated using the compounds of theinvention or a pharmaceutically acceptable salt thereof include, but arenot limited to, juvenile diabetes, diabetes mellitus, diabetes type I,or diabetes type II. In a further embodiment, protein glycosylation isnot affected by the administration of the tetracycline compounds of theinvention. In another embodiment, the tetracycline compound of theinvention is administered in combination with standard diabetictherapies, such as, but not limited to insulin therapy.

In another embodiment, the tetracycline responsive disease or disorderis a bone mass disorder. Bone mass disorders that can be treated usingthe compounds of the invention or a pharmaceutically acceptable saltthereof include disorders where a subjects bones are disorders andstates where the formation, repair or remodeling of bone isadvantageous. For examples bone mass disorders include osteoporosis (e.g., a decrease in bone strength and density), bone fractures, boneformation associated with surgical procedures (e. g., facialreconstruction), osteogenesis imperfecta (brittle bone disease),hypophosphatasia, Paget's disease, fibrous dysplasia, osteopetrosis,myeloma bone disease, and the depletion of calcium in bone, such as thatwhich is related to primary hyperparathyroidism. Bone mass disordersinclude all states in which the formation, repair or remodeling of boneis advantageous to the subject as well as all other disorders associatedwith the bones or skeletal system of a subject which can be treated withthe tetracycline compounds of the invention. In a further embodiment,the bone mass disorders include those described in U.S. Pat. Nos.5,459,135; 5,231,017; 5,998,390; 5,770,588; RE 34,656; 5,308,839;4,925,833; 3,304,227; and 4,666,897, each of which is herebyincorporated herein by reference in its entirety.

In another embodiment, the tetracycline responsive disease or disorderis acute lung injury. Acute lung injuries that can be treated using thecompounds of the invention or a pharmaceutically acceptable salt thereofinclude adult respiratory distress syndrome (ARDS), post-pump syndrome(PPS), and trauma. Trauma includes any injury to living tissue caused byan extrinsic agent or event. Examples of trauma include, but are notlimited to, crush injuries, contact with a hard surface, or cutting orother damage to the lungs.

The tetracycline responsive disease or disorders of the invention alsoinclude chronic lung disorders. Examples of chronic lung disorders thatcan be treated using the compounds of the invention or apharmaceutically acceptable salt thereof include, but are not limited,to asthma, cystic fibrosis, chronic obstructive pulmonary disease(COPD), and emphysema. In a further embodiment, the acute and/or chroniclung disorders that can be treated using the compounds of the inventionor a pharmaceutically acceptable salt thereof include those described inU.S. Pat. Nos. 5,977,091; 6,043,231; 5,523,297; and 5,773,430, each ofwhich is hereby incorporated herein by reference in its entirety.

In yet another embodiment, the tetracycline responsive disease ordisorder is ischemia, stroke, or ischemic stroke.

In a further embodiment, the tetracycline compounds of the invention ora pharmaceutically acceptable salt thereof can be used to treat suchdisorders as described above and in U.S. Pat. Nos. 6,231,894; 5,773,430;5,919,775 and 5,789,395, incorporated herein by reference.

In another embodiment, the tetracycline responsive disease or disorderis a skin wound. The invention also provides a method for improving thehealing response of the epithelialized tissue (e.g., skin, mucosae) toacute traumatic injury (e.g., cut, burn, scrape, etc.). The methodincludes using a tetracycline compound of the invention or apharmaceutically acceptable salt thereof to improve the capacity of theepithelialized tissue to heal acute wounds. The method may increase therate of collagen accumulation of the healing tissue. The method may alsodecrease the proteolytic activity in the epithelialized tissue bydecreasing the collagenolytic and/or gellatinolytic activity of MMPs. Ina further embodiment, the tetracycline compound of the invention or apharmaceutically acceptable salt thereof is administered to the surfaceof the skin (e. g., topically). In a further embodiment, thetetracycline compound of the invention or a pharmaceutically acceptablesalt thereof is used to treat a skin wound, and other such disorders asdescribed in, for example, U.S. Pat. Nos. 5,827,840; 4,704,383;4,935,412; 5,258,371; 5,308,839, 5,459,135; 5,532,227; and 6,015,804;each of which is incorporated herein by reference in its entirety.

In yet another embodiment, the tetracycline responsive disease ordisorder is an aortic or vascular aneurysm in vascular tissue of asubject (e.g., a subject having or at risk of having an aortic orvascular aneurysm, etc.). The tetracycline compound or apharmaceutically acceptable salt thereof may be effective to reduce thesize of the vascular aneurysm or it may be administered to the subjectprior to the onset of the vascular aneurysm such that the aneurysm isprevented. In one embodiment, the vascular tissue is an artery, e.g.,the aorta, e.g., the abdominal aorta. In a further embodiment, thetetracycline compounds of the invention are used to treat disordersdescribed in U.S. Pat. Nos. 6,043,225 and 5,834,449, incorporated hereinby reference in their entirety.

The compounds of the invention or a pharmaceutically acceptable saltthereof can be used alone or in combination with one or more therapeuticagent in the methods of the invention disclosed herein.

The language “in combination with” another therapeutic agent ortreatment includes co-administration of the tetracycline compound andwith the other therapeutic agent or treatment as either a singlecombination dosage form or as multiple, separate dosage forms,administration of the tetracycline compound first, followed by the othertherapeutic agent or treatment and administration of the othertherapeutic agent or treatment first, followed by the tetracyclinecompound.

The other therapeutic agent may be any agent that is known in the art totreat, prevent, or reduce the symptoms of a tetracycline-responsivedisease or disorder. The choice of additional therapeutic agent(s) isbased upon the particular tetracycline-responsive disease or disorderbeing treated. Such choice is within the knowledge of a treatingphysician. Furthermore, the other therapeutic agent may be any agent ofbenefit to the patient when administered in combination with theadministration of a tetracycline compound.

As used herein, the term “subject” means a mammal in need of treatmentor prevention, e.g., companion animals (e.g., dogs, cats, and the like),farm animals (e.g., cows, pigs, horses, sheep, goats and the like) andlaboratory animals (e.g., rats, mice, guinea pigs and the like).Typically, the subject is a human in need of the specified treatment.

As used herein, the term “treating” or ‘treatment” refers to obtainingdesired pharmacological and/or physiological effect. The effect caninclude achieving, partially or substantially, one or more of thefollowing results: partially or totally reducing the extent of thedisease, disorder or syndrome; ameliorating or improving a clinicalsymptom or indicator associated with the disorder; delaying, inhibitingor decreasing the likelihood of the progression of the disease, disorderor syndrome.

As used herein, “preventing” or “prevention” refers to reducing thelikelihood of the onset or development of disease, disorder or syndrome.

“Effective amount” means that amount of active compound agent thatelicits the desired biological response in a subject. In one embodiment,the effective amount of a compound of the invention is from about 0.01mg/kg/day to about 1000 mg/kg/day, from about 0.1 mg/kg/day to about 100mg/kg/day, or from about 0.5 mg/kg/day to about 50 mg/kg/day.

The invention further includes the process for making the compositioncomprising mixing one or more of the present compounds and an optionalpharmaceutically acceptable carrier; and includes those compositionsresulting from such a process, which process includes conventionalpharmaceutical techniques.

The compositions of the invention include ocular, oral, nasal,transdermal, topical with or without occlusion, intravenous (both bolusand infusion), inhalable, and injection (intraperitoneally,subcutaneously, intramuscularly, intratumorally, or parenterally)formulations. The composition may be in a dosage unit such as a tablet,pill, capsule, powder, granule, liposome, ion exchange resin, sterileocular solution, or ocular delivery device (such as a contact lens andthe like facilitating immediate release, timed release, or sustainedrelease), parenteral solution or suspension, metered aerosol or liquidspray, drop, ampoule, auto-injector device, or suppository; foradministration ocularly, orally, intranasally, sublingually,parenterally, or rectally, or by inhalation or insufflation.

Compositions of the invention suitable for oral administration includesolid forms such as pills, tablets, caplets, capsules (each includingimmediate release, timed release, and sustained release formulations),granules and powders; and, liquid forms such as solutions, syrups,elixirs, emulsions, and suspensions. Forms useful for ocularadministration include sterile solutions or ocular delivery devices.Forms useful for parenteral administration include sterile solutions,emulsions, and suspensions.

The compositions of the invention may be administered in a form suitablefor once-weekly or once-monthly administration. For example, aninsoluble salt of the active compound may be adapted to provide a depotpreparation for intramuscular injection (e.g., a decanoate salt) or toprovide a solution for ophthalmic administration.

The dosage form containing the composition of the invention contains aneffective amount of the active ingredient necessary to provide atherapeutic effect. The composition may contain from about 5,000 mg toabout 0.5 mg (preferably, from about 1,000 mg to about 0.5 mg) of acompound of the invention or salt form thereof and may be constitutedinto any form suitable for the selected mode of administration. Thecomposition may be administered about 1 to about 5 times per day. Dailyadministration or post-periodic dosing may be employed.

For oral administration, the composition is preferably in the form of atablet or capsule containing, e.g., 500 to 0.5 milligrams of the activecompound. Dosages will vary depending on factors associated with theparticular patient being treated (e.g., age, weight, diet, and time ofadministration), the severity of the condition being treated, thecompound being employed, the mode of administration, and the strength ofthe preparation.

The oral composition is preferably formulated as a homogeneouscomposition, wherein the active ingredient is dispersed evenlythroughout the mixture, which may be readily subdivided into dosageunits containing equal amounts of a compound of the invention.Preferably, the compositions are prepared by mixing a compound of theinvention (or pharmaceutically acceptable salt thereof) with one or moreoptionally present pharmaceutical carriers (such as a starch, sugar,diluent, granulating agent, lubricant, glidant, binding agent, anddisintegrating agent), one or more optionally present inertpharmaceutical excipients (such as water, glycols, oils, alcohols,flavoring agents, preservatives, coloring agents, and syrup), one ormore optionally present conventional tableting ingredients (such as cornstarch, lactose, sucrose, sorbitol, talc, stearic acid, magnesiumstearate, dicalcium phosphate, and any of a variety of gums), and anoptional diluent (such as water).

Binder agents include starch, gelatin, natural sugars (e.g., glucose andbeta-lactose), corn sweeteners and natural and synthetic gums (e.g.,acacia and tragacanth). Disintegrating agents include starch, methylcellulose, agar, and bentonite.

Tablets and capsules represent an advantageous oral dosage unit form.Tablets may be sugarcoated or filmcoated using standard techniques.Tablets may also be coated or otherwise compounded to provide aprolonged, control-release therapeutic effect. The dosage form maycomprise an inner dosage and an outer dosage component, wherein theouter component is in the form of an envelope over the inner component.The two components may further be separated by a layer which resistsdisintegration in the stomach (such as an enteric layer) and permits theinner component to pass intact into the duodenum or a layer which delaysor sustains release. A variety of enteric and non-enteric layer orcoating materials (such as polymeric acids, shellacs, acetyl alcohol,and cellulose acetate or combinations thereof) may be used.

Compounds of the invention may also be administered via a slow releasecomposition; wherein the composition includes a compound of theinvention and a biodegradable slow release carrier (e.g., a polymericcarrier) or a pharmaceutically acceptable non-biodegradable slow releasecarrier (e.g., an ion exchange carrier).

Biodegradable and non-biodegradable slow release carriers are well knownin the art. Biodegradable carriers are used to form particles ormatrices which retain an active agent(s) and which slowlydegrade/dissolve in a suitable environment (e.g., aqueous, acidic, basicand the like) to release the agent. Such particles degrade/dissolve inbody fluids to release the active compound(s) therein. The particles arepreferably nanoparticles or nanoemulsions (e.g., in the range of about 1to 500 nm in diameter, preferably about 50-200 nm in diameter, and mostpreferably about 100 nm in diameter). In a process for preparing a slowrelease composition, a slow release carrier and a compound of theinvention are first dissolved or dispersed in an organic solvent. Theresulting mixture is added into an aqueous solution containing anoptional surface-active agent(s) to produce an emulsion. The organicsolvent is then evaporated from the emulsion to provide a colloidalsuspension of particles containing the slow release carrier and thecompound of the invention.

The compound disclosed herein may be incorporated for administrationorally or by injection in a liquid form such as aqueous solutions,suitably flavored syrups, aqueous or oil suspensions, flavored emulsionswith edible oils such as cottonseed oil, sesame oil, coconut oil orpeanut oil and the like, or in elixirs or similar pharmaceuticalvehicles. Suitable dispersing or suspending agents for aqueoussuspensions, include synthetic and natural gums such as tragacanth,acacia, alginate, dextran, sodium carboxymethylcellulose,methylcellulose, polyvinyl-pyrrolidone, and gelatin. The liquid forms insuitably flavored suspending or dispersing agents may also includesynthetic and natural gums. For parenteral administration, sterilesuspensions and solutions are desired. Isotonic preparations, whichgenerally contain suitable preservatives, are employed when intravenousadministration is desired.

The compounds may be administered parenterally via injection. Aparenteral formulation may consist of the active ingredient dissolved inor mixed with an appropriate inert liquid carrier. Acceptable liquidcarriers usually comprise aqueous solvents and other optionalingredients for aiding solubility or preservation. Such aqueous solventsinclude sterile water, Ringer's solution, or an isotonic aqueous salinesolution. Other optional ingredients include vegetable oils (such aspeanut oil, cottonseed oil, and sesame oil), and organic solvents (suchas solketal, glycerol, and formyl). A sterile, non-volatile oil may beemployed as a solvent or suspending agent. The parenteral formulation isprepared by dissolving or suspending the active ingredient in the liquidcarrier whereby the final dosage unit contains from 0.005 to 10% byweight of the active ingredient. Other additives include preservatives,isotonizers, solubilizers, stabilizers, and pain-soothing agents.Injectable suspensions may also be prepared, in which case appropriateliquid carriers, suspending agents and the like may be employed.

Compounds of the invention may be administered intranasally using asuitable intranasal vehicle.

In another embodiment, the compounds of this invention may beadministered directly to the lungs by inhalation.

Compounds of the invention may also be administered topically orenhanced by using a suitable topical transdermal vehicle or atransdermal patch.

For ocular administration, the composition is preferably in the form ofan ophthalmic composition. The ophthalmic compositions are preferablyformulated as eye-drop formulations and filled in appropriate containersto facilitate administration to the eye, for example a dropper fittedwith a suitable pipette. Preferably, the compositions are sterile andaqueous based, using purified water. In addition to the compound of theinvention, an ophthalmic composition may contain one or more of: a) asurfactant such as a polyoxyethylene fatty acid ester; b) a thickeningagents such as cellulose, cellulose derivatives, carboxyvinyl polymers,polyvinyl polymers, and polyvinylpyrrolidones, typically at aconcentration n the range of about 0.05 to about 5.0% (wt/vol); c) (asan alternative to or in addition to storing the composition in acontainer containing nitrogen and optionally including a free oxygenabsorber such as Fe), an anti-oxidant such as butylated hydroxyanisol,ascorbic acid, sodium thiosulfate, or butylated hydroxytoluene at aconcentration of about 0.00005 to about 0.1% (wt/vol); d) ethanol at aconcentration of about 0.01 to 0.5% (wt/vol); and e) other excipientssuch as an isotonic agent, buffer, preservative, and/or pH-controllingagent. The pH of the ophthalmic composition is desirably within therange of 4 to 8.

In certain embodiments, the composition of this invention includes oneor more additional agents. The other therapeutic agent may be ay agentthat is capable of treating, preventing or reducing the symptoms of atetracycline-responsive disease or disorder. Alternatively, the othertherapeutic agent may be any agent of benefit to a patient whenadministered in combination with the tetracycline compound in thisinvention.

While this invention has been particularly shown and described withreferences to example embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims.

EXEMPLIFICATION

The following abbreviations and the terms have the indicated meanings:

Abbreviation/Term Meaning Ac acetyl AIBN2,2′-azobis(2-methylpropionitrile) aq aqueous Bn benzyl brine saturatedaqueous sodium chloride Boc tert-butoxy carbonyl or t-butoxy carbonyl(Boc)₂O di-tert-butyl dicarbonate BBr₃ boron tribromide Bu butyl Cbzbenzyloxycarbonyl CH₂Cl₂ methylene chloride CH₃CN or MeCN acetonitrileCy tricyclohexylphosphine dba dibenzylideneacetone DIBAL-Hdiisobutylaluminum hydride DIEA N,N-diisopropylethylamine DMAP4-(dimethylamino)pyridine DME 1,2-dimethoxyethane DMFN,N-dimethylformamide DMPU 1,3-dimethyl-3,4-5,6-tetrahydro-2(1H)-pyrimidone DMSO dimethyl sulfoxide EDCN-(3-dimethylaminopropyl)-N′-ethylcarbodiimide ESI electrosprayionization equiv. equivalent Et ethyl Et₂O ethyl ether EtOAc ethylacetate h, hr hour HCl hydrochloric acid KHPO₄ potassiumhydrogenphosphate HPLC high performance liquid chromatography HOBt1-hydroxybenzotriazole i iso IBX 2-iodoxybenzoic acid LDA lithiumdiisopropylamide LHMDS lithium bis(trimethylsilyl)amide LTMP lithium2,2,6,6-tetramethylpiperidide Me methyl MeOH methanol MeI methyl iodidemin minute Ms methanesulfonyl MS mass spectrum MTBE methyl tert-butylether MW molecular weight NaHCO₃ sodium bicarbonate NaOH sodiumhydroxide Na₂SO₄ sodium sulfate NBS N-bromosuccinimide NCSN-chlorosuccinimide NMR nuclear magnetic resonance spectrometry Phphenyl Pr propyl s secondary t tertiary RP reverse phase TMEDAtetramethylethylenediamine TBS tert-butyldimethylsilyl TEA triethylamineTf trifluoromathanesulfonyl TFA trifluoroacetic acid TFAAtrifluoroacetic anhydride THF tetrahydrofuran TLC thin layerchromatography Ts para-toluenesulfonyl TsOH para-toluenesulfonic acidXantphos 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene

Example 1. Synthesis of Compounds of Structural Formula (I)

The compounds of the invention can be prepared according the syntheticscheme shown in Scheme 1.

Specific conditions for the reactions depicted in Scheme 1 are providedin the following examples.

To a THF solution of 5-fluoro-2-methoxybenzoic acid (500 mg, 2.94 mmol,Aldrich 523097) cooled at −78° C. was added a THF solution of s-BuLi(4.60 mL, 1.40 M, 6.44 mmol, 2.2 equiv) and TMEDA (0.97 mL, 6.47 mmol,2.2 equiv). The reaction was stirred at −78° C. for 2 h. Mel (1.10 mL,17.64 mmol, 6.0 equiv) was added to the reaction mixture dropwise. Thereaction was allowed to warm to 25° C. over 1 h and stirred at 25° C.for 1 h. NaOH (6 N, 20 mL) was added. The resulting mixture wasextracted with t-butylmethyl ether (20 mL×2). The aqueous layer wasacidified with HCl (6 N) to pH 1 and extracted with EtOAc (20 mL×4). Thecombined EtOAc extracts were dried (Na₂SO₄) and concentrated to give 510mg of crude product 1: ¹H NMR (400 MHz, CDCl₃) δ 7.06 (dd, J=9.8, 8.5Hz, 1H), 6.75 (dd, J=9.8, 3.7 Hz, 1H), 3.86 (s, 3H), 2.34 (d, J=2.4 Hz,3H); MS (ESI) m/z 185.12 (M+H).

Oxalyl chloride (0.95 mL, 11.10 mmol, 5.5 equiv) was added to CH₂Cl₂solution (15 mL, anhydrous) of 1 (510 mg, 2.00 mmol). DMF (0.1 mL) wasadded to the resulting mixture. The reaction was stirred at 25° C. for 1h and concentrated. The resulting solid was re-dissolved in 15 mL ofanhydrous CH₂Cl₂. Phenol (520 mg, 5.50 mmol, 2.8 equiv), DMAP (670 mg,5.6 mmol, 2.8 equiv), and triethylamine (1.90 mL, 13.90 mmol, 7.0 equiv)were added to the reaction mixture. The reaction was stirred at 25° C.for 12 h and concentrated. EtOAc and H₂O were added to the residue. Theorganic layer was washed with NaOH (1 N), H₂O, and brine, dried(Na₂SO₄), and concentrated. Flash chromatography on silica gel (40:1hexanes/EtOAc) yielded 400 mg of compound 2 (52% for 2 steps): ¹H NMR(400 MHz, CDCl₃) δ 7.47-7.41 (m, 2H), 7.31-7.24 (m, 3H), 7.08 (dd,J=9.2, 9.2 Hz, 1H), 6.77 (dd, J=9.2, 3.7 Hz, 1H), 3.88 (s, 3H), 2.36 (d,J=2.3 Hz, 3H); MS (ESI) m/z 261.12 (M+H).

BBr₃ (1.85 mL, 1 M, 1.85 mmol, 1.2 equiv) was added to a CH₂Cl₂ solution(8 mL) of 2 (400 mg, 1.54 mmol) at −78° C. The reaction was stirred from−78° C. to 25° C. for 1.5 h, quenched with saturated NaHCO₃ andconcentrated. EtOAc and H₂O were added to the reaction mixture. Theaqueous layer was extracted with EtOAc. The combined EtOAc extracts weredried (Na₂SO₄) and concentrated to yield 360 mg of crude 3: ¹H NMR (400MHz, CDCl₃) δ 10.66 (s, 1H), 7.50-7.44 (m, 2H), 7.36-7.31 (m, 1H),7.26-7.18 (m, 3H), 6.86 (dd, J=9.3, 4.9 Hz, 1H), 2.60 (d, J=2.4 Hz, 3H);MS (ESI) m/z 245.11 (M−H).

Boc₂O (350 mg, 1.60 mmol, 1.1 equiv) and DMAP (20 mg, 0.16 mmol, 0.1equiv) were added to a CH₂Cl₂ solution of crude 3 (360 mg). The reactionwas stirred at 25° C. for 1.5 h and concentrated. Flash chromatographyon silica gel (35:1 hexanes/EtOAc) yielded 400 mg of compound 4 (94% for2 steps): ¹H NMR (400 MHz, CDCl₃) δ 7.46-7.41 (m, 2H), 7.31-7.23 (m, 3H)7.18 (dd, J=8.8, 8.7 Hz, 1H), 7.10 (dd, J=8.8, 4.4 Hz, 1H), 2.41 (d,J=2.3 Hz, 3H), 1.44 (s, 9H); MS (ESI) m/z 345.18 (M−H).

A THF solution (6 mL) of 4 (487 mg, 1.40 mmol, 2.0 equiv) was added to aTHF solution (5 mL) of LDA (6.30 mL, 10% wt, 4.20 mmol, 6.0 equiv) andTMEDA (1.70 mL, 11.20 mmol, 16.0 equiv) at −78° C. The reaction wasstirred at −78° C. for 5 min. A THF solution of enone (339 mg, 0.70mmol, 1.0 equiv) was added to the reaction mixture dropwise. Thereaction was stirred from −78° C. to 25° C. for 1 h, quenched withsaturated NH₄Cl, and extracted with EtOAc. The combined EtOAc extractswere dried (Na₂SO₄) and concentrated to yield the crude product.Preparative reverse phase HPLC purification on a Waters Autopurificationsystem using a Sunfire Prep C18 OBD column [5 μm, 19×50 mm; flow rate,20 mL/min; Solvent A: H₂O with 0.1% HCO₂H; Solvent B: CH₃CN with 0.1%HCO₂H; injection volume: 4.0 mL (CH₃CN); gradient: 80→100% B over 15min; mass-directed fraction collection]. Fractions with the desired MW,eluting at 6.3-8.0 min, were collected and concentrated on a RotaVap atrt to remove most of the acetonitrile. The resulting mostly aqueoussolution was extracted with EtOAc. The combined EtOAc extracts weredried (Na₂SO₄) and concentrated to give 185 mg of pure 5 (35%): ¹H NMR(400 MHz, CDCl₃) δ 15.67 (s, 1H), 7.51-7.46 (m, 2H), 7.39-7.29 (m, 3H),7.21 (dd, J=8.9, 8.9 Hz, 1H), 7.03 (dd, J=8.9, 4.0 Hz, 1H), 5.34 (s,2H), 3.93 (d, J=10.4 Hz, 1H), 3.30-3.21 (m, 1H), 3.10-3.00 (m, 1H),2.57-2.41 (m, 3H), 2.48 (s, 6H), 2.17-2.12 (m, 1H), 1.53 (s, 9H), 0.82(s, 9H), 0.26 (s, 3H), 0.12 (s, 3H); MS (ESI) m/z 735.45 (M+H).

Aqueous HF (3 mL, 48%) and TFA (4 μL) were added to a CH₃CN solution (7mL) of 5 (210 mg, 0.29 mmol) in a polypropylene tube at 25° C. Thereaction was stirred at 25° C. for 18 h. The resulting mixture waspoured into an aqueous solution of K₂HPO₄ (21 g, dissolved in 150 mLwater). The mixture was extracted with EtOAc. The combined EtOAcextracts were dried (Na₂SO₄) and concentrated to yield 180 mg of crude6: ¹H NMR (400 MHz, CDCl₃) δ 14.64 (s, 1H), 11.47 (s, 1H), 7.49-7.45 (m,2H), 7.39-7.32 (m, 3H), 7.14 (dd, J=9.2, 8.8 Hz, 1H), 6.77 (dd, J=9.2,4.3 Hz, 1H), 5.36 (s, 2H), 3.68 (d, J=3.7 Hz, 1H), 3.09 (dd, J=15.6, 4.6Hz, 1H), 3.02-2.92 (m, 1H), 2.84-2.79 (m, 1H), 2.49 (s, 6H), 2.34-2.22(m, 1H), 2.09-2.02 (m, 1H), 1.55-1.44 (m, 1H); MS (ESI) m/z 521.30(M+H).

Palladium on carbon (35 mg, 10 wt %) was added to a MeOH/dioxanesolution (4 mL/4 mL) of crude 6 (180 mg). The reaction was purged withhydrogen and stirred under H₂ (balloon) at 25° C. for 1 h. The reactionmixture was filtered through a small Celite plug. The filtrate wasconcentrated to yield the crude product. Preparative reverse phase HPLCpurification on a Waters Autopurification system using a PhenomenexPolymerx 10μ RP-1 100 A column [10 μm, 150×21.20 mm; flow rate, 20mL/min; Solvent A: 0.05 N HCl/water; Solvent B: CH₃CN; injection volume:4.0 mL (0.05 N HCl/water); gradient: 0→100% B over 15 min; mass-directedfraction collection]. Fractions with the desired MW, eluting at 6.4-8.2min, were collected and freeze-dried to yield 51 mg of compound 7 (41%for 2 steps): ¹H NMR (400 MHz, CD₃OD) δ 7.26 (dd, J=9.2, 9.2 Hz, 1H),6.80 (dd, J=9.2, 4.3 Hz, 1H), 4.09 (br s, 1H), 3.14 (dd, J=15.0, 4.6 Hz,1H), 3.04 (s, 3H), 2.96 (s, 3H), 3.09-2.91 (m, 2H), 2.31-2.18 (m, 2H),1.68-1.56 (m, 1H); MS (ESI) m/z 433.28 (M+H).

A mixture of HNO₃ (8.5 μL, 69%) and H₂SO₄ (0.5 mL) was added to a H₂SO₄solution (1 mL) of 7 (51 mg, 0.12 mmol) at 0° C. The reaction wasstirred at 0° C. for 30 min. The resulting mixture was added dropwise tovigorously stirred diethyl ether (60 mL). The suspension was filteredthrough a small Celite pad and washed several times with more diethylether. The Celite pad was then eluted with MeOH until the eluent becamecolorless. The yellow MeOH eluent was collected and concentrated underreduced pressure to afford crude 8: ¹H NMR (400 MHz, CD₃OD) δ 8.03 (d,J=8.5 Hz, 1H), 4.09 (br s, 1H), 3.50-2.97 (m, 3H), 3.04 (s, 3H), 2.96(s, 3H), 2.46-2.36 (m, 1H), 2.29-2.20 (m, 1H), 1.71-1.59 (m, 1H); MS(ESI) m/z 478.20 (M+H).

Palladium on carbon (12 mg, 10 wt %) was added to a MeOH solution (4 mL)of crude 8. The reaction was purged with hydrogen and stirred under H₂(balloon) at 25° C. for 2 h. The catalyst was filtered off with a smallCelite pad. The filtrate was concentrated to yield crude 9. Preparativereverse phase HPLC purification on a Waters Autopurification systemusing a Phenomenex Polymerx 10μ RP-1 100 A column [10 μm, 150×21.20 mm;flow rate, 20 mL/min; Solvent A: 0.05 N HCl; Solvent B: CH₃CN; injectionvolume: 4.0 mL (0.05 N HCl/water); gradient: 0→100% B over 15 min;mass-directed fraction collection]. Fractions with the desired MW,eluting at 5.0-6.6 min, were collected and freeze-dried to yield 43 mgof pure 9 (81% for 2 steps): ¹H NMR (400 MHz, CD₃OD) δ 7.43 (d, J=8.5Hz, 1H), 4.11 (br s, 1H), 3.22-3.16 (m, 1H), 3.15-3.08 (m, 1H),3.06-2.95 (m, 1H), 3.04 (s, 3H), 2.96 (s, 3H), 2.40-2.31 (m, 1H),2.28-2.21 (m, 1H), 1.71-1.59 (m, 1H); MS (ESI) m/z 448.24 (M+H).

2-t-Butylaminoacetylchloride hydrochloride (4.2 mg, 0.022 mmol, 2.0equiv) was added to a DMF solution (0.1 mL) of 9 (5 mg, 0.011 mmol) at25° C. The reaction was stirred at 25° C. for 30 min. The reactionmixture was diluted with 0.05 N HCl (2 mL) and injected into a WatersAutopurification system equipped with a Phenomenex Polymerx 10μ RP-1 100A column [10 μm, 150×21.20 mm; flow rate, 20 mL/min; Solvent A: 0.05 NHCl; Solvent B: CH₃CN; gradient: 0→100% B over 20 min; mass-directedfraction collection]. Fractions with the desired MW, eluting at 6.4-7.0min, were collected and freeze-dried to yield 3.9 mg of pure 11 (62%):¹H NMR (400 MHz, CD₃OD) δ 8.25 (d, J=11.0 Hz, 1H), 4.11 (br s, 1H), 4.09(s, 2H), 3.22-2.86 (m, 3H), 3.05 (s, 3H), 2.97 (s, 3H), 2.33-2.20 (m,2H), 1.69-1.57 (m, 1H), 1.42 (s, 9H); MS (ESI) m/z 561.39 (M+H).

Anhydrous Na₂CO₃ (16 mg, 0.15 mmol, 5.5 equiv) was added to an anhydrousDMPU/acetonitrile (150 μL/50 μL) solution of 9 (12 mg, 0.027 mmol).Bromoacetyl bromide (2.8 μL, 0.032 mmol, 1.2 equiv) was added to themixture. The reaction was stirred at 25° C. for 10 min. LC/MS analysisindicated complete formation of intermediate 10. Azetidine (36 μL, 0.54mmol, 20 equiv) was added to the reaction mixture. The reaction wasstirred at 25° C. for 2 h. The reaction mixture was concentrated andacidified with HCl (0.5 N in MeOH, 0.7 mL). The resulting mixture wasadded dropwise to vigorously stirred diethyl ether (10 mL). Thesuspension was filtered through a small Celite pad and washed severaltimes with more diethyl ether. The Celite pad was then eluted with MeOHuntil the eluent became colorless. The yellow MeOH eluent was collectedand concentrated under reduced pressure to afford crude 32. Preparativereverse phase HPLC purification on a Waters Autopurification systemusing a Phenomenex Polymerx 10μ RP-1 100 A column [10 pm, 150×21.20 mm;flow rate, 20 mL/min; Solvent A: 0.05 N HCl; Solvent B: CH₃CN; injectionvolume: 2.0 mL (0.05 N HCl/water); gradient: 10→20% B over 30 min;mass-directed fraction collection]. Fractions with the desired MW,eluting at 10.8-12.5 min, were collected and freeze-dried to yield 2.0mg of pure 32: ¹H NMR (400 MHz, CD₃OD) δ 8.18 (d, J=11.0 Hz, 1H),4.41-4.31 (m, 2H), 4.32 (s, 2H), 4.24-4.13 (m, 2), 4.08 (br s, 1H),3.18-2.86 (m, 3H), 3.03 (s, 3H), 2.95 (s, 3H), 2.71-2.57 (m, 1H),2.54-2.42 (m, 1H), 2.33-2.16 (m, 2H), 1.69-1.57 (m, 1H); MS (ESI) m/z545.20 (M+H).

Compounds 12-31 and Compounds 33-46 are prepared similarly to Compounds11 or 32, substituting the appropriate acyl halide for2-t-Butylaminoacetylchloride in the synthesis of Compound 11 or cyclicamine for azetidine in the synthesis of Compound 32.

¹H NMR (400 MHz, CD₃OD) δ 8.23 (d, J=11.2 Hz, 1H), 4.08 (s, 3H),3.17-2.97 (m, 11H), 2.31 (dd, J=14.8, 14.8 Hz, 1H), 2.24 (ddd, J=14.0,5.2, 2.8 Hz, 1H), 1.79-1.72 (m, 2H), 1.66 (ddd, J=13.6, 13.6, 13.6 Hz,1H), 1.05 (t, J=7.2 Hz, 3H); MS (ESI) m/z 547.2 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.23 (d, J=11.2 Hz, 1H), 4.08 (s, 3H),3.16-2.97 (m, 11H), 2.30 (dd, J=14.8, 14.8 Hz, 1H), 2.24 (ddd, J=14.4,5.2, 2.8 Hz, 1H), 1.75-1.69 (m, 2H), 1.66 (ddd, J=13.6, 13.6, 13.6 Hz,1H), 1.49-1.41 (m, 2H), 1.01 (t, J=7.2 Hz, 3H); MS (ESI) m/z 561.2(M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.21 (d, J=11.2 Hz, 1H), 4.08 (s, 1H), 4.06(s, 2H), 3.16-2.96 (m, 11H), 2.28 (dd, J=14.8, 14.8 Hz, 1H), 2.22 (ddd,J=14.4, 5.2, 2.8 Hz, 1H), 1.77-1.71 (m, 2H), 1.66 (ddd, J=14.0, 14.0,14.0 Hz, 1H), 1.43-1.35 (m, 6H), 0.93 (t, J=7.2 Hz, 3H); MS (ESI) m/z589.2 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.23 (d, J=10.8 Hz, 1H), 4.09 (s, 2H), 4.07(s, 1H), 3.15-2.95 (m, 11H), 2.29 (dd, J=14.4, 14.4 Hz, 1H), 2.25 (ddd,J=14.4, 5.2, 2.8 Hz, 1H), 1.66 (ddd, J=13.2, 13.2, 13.2 Hz, 1H), 1.10(s, 9H); MS (ESI) m/z 575.2 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.24 (d, J=11.0 Hz, 1H), 4.08 (s, 2H),4.01-3.89 (m, 1H), 3.50-3.42 (m, 1H), 3.20-2.84 (m, 9H), 2.30 (at,J=14.7 Hz, 1H), 2.23-2.15 (m, 1H), 1.70-1.58 (m, 1H), 1.37 (d, J=6.7 Hz,6H); MS (ESI) m/z 547.25 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.24 (d, J=11.0 Hz, 1H), 4.20 (s, 2H), 4.09(br s, 1H), 3.19-3.13 (m, 1H), 3.12-2.89 (m, 2H), 2.89-2.38 (m, 1H),3.04 (s, 3H), 2.96 (s, 3H), 2.35-2.19 (m, 2H), 1.71-1.59 (m, 1H), 0.95(br s, 2), 0.94 (br s, 2); MS (ESI) m/z 545.37 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.23 (d, J=10.8 Hz, 1H), 4.09 (s, 3H),3.68-3.61 (m, 1H), 3.16-2.97 (m, 9H), 2.29 (dd, J=14.4, 14.4 Hz, 1H),2.25 (ddd, J=14.4, 5.2, 2.8 Hz, 1H), 2.20-2.12 (m, 2H), 1.98-1.91 (m,2H), 1.75-1.68 (m, 4H), 1.66 (ddd, J=13.6, 13.6, 13.6 Hz, 1H); MS (ESI)m/z 573.1 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.26 (d, J=11.0 Hz, 1H), 4.09 (br s, 3H),3.19-2.93 (m, 5H), 3.04 (s, 3H), 2.96 (s, 3H), 2.35-2.26 (m, 1H),2.25-2.18 (m, 1H), 2.14-2.02 (m, 1H), 1.71-1.59 (m, 1H), 1.07 (d, J=6.7,6 H); MS (ESI) m/z 561.24 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.24 (d, J=11.0 Hz, 1H), 4.11 (s, 2H), 4.08(br s, 1H), 3.22-2.92 (m, 5H), 3.03 (s, 3H), 2.95 (s, 3H), 2.33-2.24 (m,1H), 2.24-2.17 (m, 1H), 1.69-1.58 (m, 1H), 1.17-1.07 (m, 1H), 0.77-0.71(m, 2H), 0.46-0.40 (m, 2H); MS (ESI) m/z 559.23 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.25 (d, J=11.0 Hz, 1H), 4.12 (s, 2H), 4.09(s, 1H), 3.72-3.67 (m, 2H), 3.43 (s, 3H), 3.19-2.92 (m, 11H), 2.35-2.18(m, 2H) 1.71-1.58 (m, 1H); MS (ESI) m/z 563.23 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.25 (d, J=11.0 Hz, 1H), 4.22 (s, 2H),4.14-4.05 (m, 3H), 3.18-2.84 (m, 9H), 2.34-2.17 (m, 2H), 1.70-1.57 (m,1H); MS (ESI) m/z 587.28 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.24 (d, J=11.0 Hz, 1H), 4.24 (s, 2H), 4.09(s, 1H), 3.14-2.93 (m, 15H), 2.24-2.18 (m, 2H), 1.65 (dt, J=13.4, 11.6Hz, 1H); MS (ESI) m/z 533.17 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.23 (d, J=10.4 Hz, 1H), 4.29 (d, J=16.5 Hz,1H), 4.18 (d, J=15.9 Hz, 1H), 4.09 (s, 1H), 3.19-2.89 (m, 14H),2.36-2.17 (m, 2H), 1.70-1.58 (m, 1H), 1.38 (t, J=7.32 Hz, 3H); MS (ESI)m/z 547.25 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.21 (d, J=10.8 Hz, 1H), 4.25 (s, 2H), 4.10(s, 1H), 3.35 (t, J=7.2 Hz, 3H), 3.34 (t, J=7.2 Hz, 3H), 3.13-2.99 (m,9H), 2.31 (dd, J=14.8, 14.8 Hz, 1H), 2.27 (ddd, J=14.8, 5.2, 2.8 Hz,1H), 1.78-1.74 (m, 2H), 1.68 (ddd, J=13.6, 13.6, 13.6 Hz, 1H), 1.38 (t,J=7.2 Hz, 6H); MS (ESI) m/z 561.2 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.23 (d, J=11.2 Hz, 1H), 4.10 (s, 3H),3.16-2.96 (m, 11H), 2.31 (dd, J=14.4, 14.4 Hz, 1H), 2.24 (ddd, J=14.4,5.2, 2.8 Hz, 1H), 1.78-1.71 (m, 2H), 1.66 (ddd, J=14.0, 14.0, 14.0 Hz,1H), 1.45-1.38 (m, 4H), 0.98 (t, J=7.2 Hz, 3H); MS (ESI) m/z 575.2(M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.24 (d, J=10.8 Hz, 1H), 4.09 (s, 3H), 3.59(t, J=5.6 Hz, 2H), 3.40 (s, 3H), 3.23 (t, J=5.6 Hz, 2H), 3.15-2.94 (m,9H), 2.32 (dd, J=15.2, 15.2 Hz, 1H), 2.24 (ddd, J=14.0, 5.2, 2.8 Hz,1H), 2.08-2.02 (m, 2H), 1.66 (ddd, J=15.2, 15.2, 15.2 Hz, 1H); MS (ESI)m/z 577.2 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.24 (d, J=10.8 Hz, 1H), 4.32 (d, J=8.0 Hz,1H), 4.21 (d, J=8.0 Hz, 1H), 4.10 (s, 1H), 3.18-2.99 (m, 9H), 3.01 (s,3H), 2.33 (dd, J=14.8, 14.8 Hz, 1H), 2.29 (ddd, J=15.2, 5.2, 2.8 Hz,1H), 1.78-1.74 (m, 2H), 1.88-1.81 (m, 2H), 1.68 (ddd, J=15.6, 15.6, 15.6Hz, 1H), 1.08 (t, J=7.2 Hz, 3H); MS (ESI) m/z 561.2 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.25 (d, J=10.8 Hz, 1H), 4.10 (s, 3H),3.18-2.98 (m, 11H), 2.31 (dd, J=14.8, 14.8 Hz, 1H), 2.26 (ddd, J=14.4,5.2, 2.8 Hz, 1H), 1.78-1.74 (m, 2H), 1.66 (ddd, J=13.6, 13.6, 13.6 Hz,1H), 1.42-1.30 (m, 8H), 0.94 (t, J=6.8 Hz, 3H); MS (ESI) m/z 603.2(M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.32 (d, J=10.4 Hz, 1H), 7.38-7.34 (m, 2H),7.10-7.06 (m, 3H), 4.17 (s, 2H), 4.10 (s, 1H), 3.18-2.99 (m, 11H), 2.29(dd, J=15.6, 15.6 Hz, 1H), 2.25 (ddd, J=14.8, 5.2, 2.8 Hz, 1H), 1.66(ddd, J=14.8, 14.8, 14.8 Hz, 1H); MS (ESI) m/z 581.1 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.25 (d, J=10.8 Hz, 1H), 4.36 (d, J=8.0 Hz,1H), 4.21 (d, J=8.0 Hz, 1H), 4.10 (s, 1H), 3.68-3.61 (m, 1H), 3.18-2.98(m, 9H), 3.00 (s, 3H), 2.29 (dd, J=14.4, 14.4 Hz, 1H), 2.20-2.10 (m,3H), 1.96-1.89 (m, 2H), 1.78-1.68 (m, 4H), 1.66 (ddd, J=14.4, 14.4, 14.4Hz, 1H); MS (ESI) m/z 587.2 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.20 (d, J=11.0 Hz, 1H), 5.54-5.33 (m, 2H),4.71-4.37 (m, 4H), 4.40 (s, 2H), 4.06 (br s, 1H), 3.17-2.92 (m, 3H),2.99 (s, 6H), 2.33-2.24 (m, 1H), 2.23-2.16 (m, 1H), 1.70-1.58 (m, 1H);MS (ESI) m/z 563.20 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.22 (d, J=11.0 Hz, 1H), 4.33 (s, 2H), 4.10(s, 1H), 3.83-3.72 (m, 2H), 3.25-2.89 (m, 12H), 2.32-2.00 (m, 6H),1.69-1.56 (m, 1H); MS (ESI) m/z 559.39 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.25 (d, J=11.0 Hz, 1H), 5.54-5.31 (m, 1H),4.39-4.20 (m, 2H), 4.09-4.01 (m, 1H), 3.40-3.30 (m, 2H), 3.09-2.89 (m,12H), 2.50-2.34 (m, 2H), 2.34-2.25 (m, 1H), 2.24-2.16 (m, 1H), 1.71-1.58(m, 1H); MS (ESI) m/z 577.32 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.23 (d, J=10.4 Hz, 1H), 5.57-5.37 (m, 1H),4.47-4.33 (m, 2H), 4.15-3.87 (m, 2H), 3.72-3.40 (m, 1H), 3.17-2.83 (m,12H), 2.55-2.34 (m, 2H), 2.33-2.18 (m, 2H), 1.69-1.57 (m, 1H); MS (ESI)m/z 577.37 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.28 (d, J=10.7 Hz, 1H), 4.08 (s, 1H),4.00-3.91 (m, 2H), 3.09-2.57 (m, 18H), 3.26-3.18 (m, 3H), 2.49-2.34 (m,2H), 2.35-2.06 (m, 2H), 1.72-1.59 (m, 1H); MS (ESI) m/z 602.37 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.24 (d, J=11.0 Hz, 1H), 4.46-4.32 (m, 2H),4.26-4.16 (m, 1H), 4.08 (s, 2H), 4.00-3.72 (m, 2H), 3.18-2.91 (m, 16H),2.68-2.56 (m, 1H), 2.51-2.39 (m, 1H), 2.34-2.24 (m, 1H), 2.23-2.17 (m,1H), 1.70-1.57 (m, 1H); MS (ESI) m/z 602.37 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.23 (d, J=11.0 Hz, 1H), 4.62-4.54 (m, 1H),4.48-4.24 (m, 2H), 4.08 (s, 1H), 3.99-3.69 (m, 3H), 3.50-3.40 (m, 1H),3.17-2.90 (m, 9H), 2.44-2.11 (m, 4H), 2.10-2.00 (m, 1H), 1.69-1.56 (m,1H); MS (ESI) m/z 575.27 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.23 (d, J=11.0 Hz, 1H), 4.62-4.54 (m, 1H),4.50-4.38 (m, 1H), 4.37-4.27 (m, 1H), 4.10 (s, 1H), 3.99-3.70 (m, 3H),3.50-3.40 (m, 1H), 3.24-2.84 (m, 9H), 2.40-2.11 (m, 4H), 2.10-2.01 (m,1H), 1.70-1.57 (m, 1H); MS (ESI) m/z 575.33 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.25 (d, J=11.0 Hz, 1H), 4.54 (d, J=16.5 Hz,1H), 4.26 (d, J=15.9 Hz, 1H), 4.09 (s, 1H), 3.95-3.81 (m, 2H), 3.81-3.75(m, 1H), 3.69-3.62 (m, 1H), 3.35 (s, 3H), 3.23-2.92 (m, 9H), 2.35-2.04(m, 6H), 1.91-1.80 (m, 1H), 1.71-1.59 (m, 1H); MS (ESI) m/z 603.35(M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.24 (d, J=11.0 Hz, 1H), 4.55 (d, J=16.5 Hz,1H), 4.27 (d, J=16.5 Hz, 1H), 4.10 (s, 1H), 3.95-3.82 (m, 2H), 3.81-3.75(m, 1H), 3.70-3.63 (m, 1H), 3.38 (s, 3H), 3.20-2.92 (m, 9H), 2.35-2.02(m, 6H), 1.92-1.80 (m, 1H), 1.70-1.58 (m, 1H); MS (ESI) m/z 603.41(M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.22 (d, J=11.0 Hz, 1H), 4.19 (s, 2H), 4.09(s, 1H), 3.65-3.58 (m, 2H), 3.19-2.92 (m, 10H), 2.34-2.18 (m, 2H),2.02-1.79 (m, 6H), 1.69-1.50 (m, 2H); MS (ESI) m/z 573.35 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.24 (d, J=11.0 Hz, 1H), 4.28 (s, 2H),4.03-4.00 (m, 2H), 3.94-3.81 (m, 2H), 3.68-3.55 (m, 2H), 3.20-2.88 (m,12H), 2.36-2.18 (m, 2H), 1.71-1.57 (m, 1H); MS (ESI) m/z 575.37 (M+H).

¹H NMR (400 MHz, CD₃OD, 2:1 mixture of diastereomers) δ 8.25 (d+d,J=11.0 Hz, 1H), 4.29, 4.24 (s+s, 2H), 4.08 (s+s, 1H), 4.01-3.92 (m+m,3H), 3.20-2.62 (m+m, 13H), 2.35-2.16 (m+m, 3H), 1.83-1.46 (m+m, 5H); MS(ESI) m/z 599.36 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.29 (d, J=11.0 Hz, 1H), 7.41 (s, 5H),4.50-4.37 (m, 2H), 4.05 (s, 1H), 3.95-3.81 (m, 2H), 3.40-3.37 (m, 1H),3.24-3.15 (m, 3H), 3.10-2.70 (m, 9H), 2.36-2.25 (m, 1H), 2.25-2.16 (m,1H), 1.72-1.59 (m, 1H); MS (ESI) m/z 607.34 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.15 (d, J=10.8 Hz, 1H), 4.00 (s, 1H), 3.99(s, 2H), 3.10-2.87 (m, 11H), 2.32-2.12 (m, 2H), 1.59-1.51 (m, 1H), 1.26(t, J=7.2 Hz, 3H); MS (ESI) m/z 533.1 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.15 (d, J=11.2 Hz, 1H), 4.00 (s, 1H), 3.96(s, 2H), 3.08-2.87 (m, 11H), 2.70-2.61 (m, 1H), 2.23-2.09 (m, 4H),1.97-1.75 (m, 4H), 1.59-1.51 (m, 1H); MS (ESI) m/z 572.2 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.26 (d, J=10.8 Hz, 1H), 4.10 (s, 3H),3.21-2.97 (m, 11H), 2.35-2.20 (m, 2H), 2.15-2.05 (m, 1H), 1.98-1.82 (m,2H), 1.77-1.61 (m, 5H), 1.35-1.26 (m, 2H); MS (ESI) m/z 587.1 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.26 (d, J=10.8 Hz, 1H), 4.16 (s, 1H), 4.14(s, 2H), 3.20-2.95 (m, 11H), 2.32-2.20 (m, 2H), 1.88-1.59 (m, 6H),1.39-1.21 (m, 4H), 1.12-1.02 (m, 2H); MS (ESI) m/z 601.1 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.14 (d, J=10.8 Hz, 1H), 4.00 (s, 1H), 3.88(s, 2H), 3.77-3.73 (m, 1H), 3.09-2.87 (m, 9H), 2.29-2.10 (m, 6H),1.88-1.81 (m, 2H), 1.59-1.50 (m, 1H); MS (ESI) m/z 559.1 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.24 (d, J=10.8 Hz, 1H), 4.10 (s, 3H),3.17-2.97 (m, 9H), 2.32-2.09 (m, 4H), 1.92-1.85 (m, 2H), 1.75-1.63 (m,2H), 1.43-1.26 (m, 6H); MS (ESI) m/z 587.2 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.20 (d, J=11.2 Hz, 1H), 8.16 (d, J=2.4 Hz,1H), 8.06 (d, J=5.2 Hz, 1H), 7.85-7.78 (m, 2H), 4.27 (s, 2H), 4.11 (s,1H), 3.18-2.98 (m, 9H), 2.32-2.21 (m, 2H), 1.70-1.60 (m, 1H); MS (ESI)m/z 582.2 (M+H)

¹H NMR (400 MHz, CD₃OD) δ 8.24 (d, J=11.0 Hz, 1H), 4.31 (s, 2H), 4.11(s, 1H), 3.22-2.88 (m, 9H), 2.36-2.16 (m, 2H), 1.70-1.56 (m, 1H), 1.44(s, 9H); MS (ESI) m/z 577.41 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.15 (d, J=10.8 Hz, 1H), 4.65 (t, J=4.8 Hz,2H), 4.08 (s, 2H), 4.00 (s, 1H), 3.45 (t, J=4.4 Hz, 1H), 3.38 (t, J=5.6Hz, 1H), 3.20-2.87 (m, 9H), 2.25-2.09 (m, 2H), 1.59-1.50 (m, 1H); MS(ESI) m/z 551.0 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.25 (d, J=11.2 Hz, 1H), 6.39 (tt, J=53.6, 3.2Hz, 1H), 4.24 (s, 2H), 4.13 (s, 1H), 3.71 (td, J=15.2, 2.8 Hz, 2H),3.19-2.91 (m, 9H), 2.33-2.24 (m, 2H), 1.70-1.60 (m, 1H); MS (ESI) m/z569.0 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.21 (d, J=10.8 Hz, 1H), 4.01 (s, 1H), 3.85(s, 2H), 3.73 (s, 3H), 3.59-3.51 (m, 1H), 3.12-2.87 (m, 9H), 2.23-2.12(m, 2H), 1.88-1.50 (m, 9H); MS (ESI) m/z 559.1 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.24 (d, J=10.8 Hz, 1H), 4.48 (s, 2H), 4.12(s, 1H), 4.10-4.07 (m, 2H), 3.93-3.86 (m, 2H), 3.19-2.90 (m, 9H),2.79-2.67 (m, 2H), 2.37-2.21 (m, 2H), 1.59-1.51 (m, 1H); MS (ESI) m/z595.0 (M+H).

Example 2. Synthesis of Compounds of Structural Formula (I), Wherein R¹and R² Taken Together with the Nitrogen to which they are Bonded Form aMonocyclic or Bicyclic Heteroaryl

The following compounds were prepared according to Scheme 2.

To a 250 mL round bottom flask was added compound 3 (14.47 g, 56.30mmol, 1.0 equiv, crude), tetrabutylammonium bromide (0.90 g, 2.80 mmol,0.05 equiv), 1,2-dichloroethane (60 mL), and water (60 mL). The clearbi-layer was cooled in a 20° C. water bath. Nitric acid (7.2 mL, 70 wt%, 112.60 mmol, 2.0 equiv) was added. After the addition, the reactiontemperature slowly rose to 26° C. The reaction was stirred at roomtemperature overnight (19 hrs). TLC (heptane/EtOAc=9.5/0.5) showed thereaction was complete. The organic layer was separated, washed withwater (60 mL×2) and brine, and dried over anhydrous sodium sulfate. Thesolvent was removed to give compound 2-1 as a brown oil, whichsolidified on standing (17.71 g, quantitative). The crude product wasused directly for the next step.

To a 250 mL round bottom flask was added compound 2-1 (17.7 g, 56.30mmol 1.0 equiv), acetone (177 mL), anhydrous potassium carbonate (15.6g, 113.00 mmol, 2.0 equiv), and potassium iodide (0.47 g, 2.80 mmol,0.05 equiv). To the stirred suspension at room temperature was addedbenzyl bromide (7.03 mL, 59.10 mmol, 1.05 equiv). The suspension wasthen heated to 56° C. for 4 hrs. TLC (heptane/EtOAc=9/1) showed thereaction was complete. The solid was removed by filtration and washedwith acetone (30 mL). The filtrated was concentrated to give a paste.The paste was partitioned between methyl t-butyl ether (MTBE, 120 mL)and water (80 mL). The organic layer was washed with water (80 mL) andbrine, dried over anhydrous sodium sulfate, and concentrated to givecompound 2-2 as a brown oil (21.09 g, 98%). The crude product was useddirectly for the next step.

To a 1 L round bottom flask was added compound 2-2 (21.08 g, 55.40 mmol,1.0 equiv) and THF (230 mL). The solution was cooled in a cold waterbath to 10° C. To another 500 mL round bottom flask containing water(230 mL), sodium hydrosulfite (Na₂S₂O₄, 56.7 g, 276.80 mmol, 5.0 equiv)was added slowly with stirring. The aqueous solution of sodiumhydrosulfite was added to the THF solution of compound 2-2. Thetemperature quickly rose from 10° C. to 20.4° C. after the addition. Theyellow suspension was stirred while the cold water bath slowly warmed upto room temperature overnight to give an orange cloudy solution. Thereaction temperature during this period was between 15° C. to 19° C. TLC(heptane/EtOAc=9/1) showed the reaction was complete. The orange cloudysolution was diluted with EtOAc (460 mL). The organic layer was washedwith water (150 mL×2) and brine, dried over anhydrous sodium sulfate,and concentrated under reduced pressure to give the crude product as abrown oil. The crude product was purified by flash silica gel columneluted with heptane/EtOAc 9/1 to yield the desired product 2-3 (15.83 g,80%, 3 steps).

To an NMP solution (50 mL) aniline 2-3 (10.02 g, 28.5 mmol, 1 equiv) wasadded allyl bromide (7.65 mL, 85.5 mmol, 3 equiv) and potassiumcarbonate (11.79 g, 85.5 mmol, 3 equiv). Potassium iodide (994.8 mg, 6mmol, 0.2 equiv) was added and the reaction mixture was placed undernitrogen and heated to 100° C. After 16 h, the reaction was cooled,diluted with water (60 mL), and extracted with EtOAc (75 mL, then 2×50mL). The combined organic extracts were washed with water (2×35 mL),were dried (Na₂SO₄), filtered, and concentrated to yield the crudeproduct. Purification via flash column chromatography on silica gel(RediSep, 125 g, gradient 1-6% EtOAc in hexanes) gave 10.97 g of pure2-4 (89%): ¹H NMR (400 MHz, CDCl₃) δ 7.42-7.30 (m, 7H), 7.42-7.20 (m,1H), 7.00 (d, J=8.5 Hz, 2H), 6.72 (d, J=11.0 HZ, 1H), 5.77-5.70 (m, 2H),5.20-5.12 (m, 6H), 3.81 (d, J=6.1 Hz, 4H), 2.26 (s, 3H); MS (ESI) m/z432.34 (M+H).

A THF solution (6.5 mL) of 2-4 (875 mg, 2.03 mmol, 1.25 equiv) was addedto a freshly-prepared solution of LDA in THF (0.051 M, 2.03 mmol, 40 mL,1.25 equiv) and TMEDA (304 μL, 2.03 mmol, 1.25 equiv) at −78° C. Thereaction was stirred at −78° C. for 15 min. A THF solution (6.5 mL) ofenone (784 mg, 1.62 mmol, 1.0 equiv) was added to the reaction mixturedropwise, followed by addition of LHMDS solution (1.0 M in THF, 2.03 mL,2.03 mmol, 1.25 equiv). The reaction was stirred from −78° C. to −10° C.for 1 h, quenched with saturated NH₄Cl (6 mL), and warmed to 25° C. Thesolution was poured into saturated NH₄Cl (20 mL) and extracted withEtOAc (2×75 mL). The combined EtOAc extracts were dried (Na₂SO₄),filtered, and concentrated to yield the crude product. Preparativereverse phase HPLC purification on a Waters Autopurification systemusing a Sunfire Prep C18 OBD column [5 μm, 19×50 mm; flow rate, 20mL/min; Solvent A: H₂O with 0.1% HCO₂H; Solvent B: CH₃CN with 0.1%HCO₂H; injection volume: 4×3.6-4.2 mL (CH₃CN); gradient: 88→100% B over12 min; mass-directed fraction collection]. Fractions with the desiredMW, eluting at 6.6-10.6 min, were collected and lyophilized to give 552mg of pure 2-5 (41%): ¹H NMR (400 MHz, CDCl₃) δ 16.22 (s, 1H), 7.49-7.47(m, 4H), 7.37-7.31 (m, 6H), 6.80 (d, J=11.0 Hz, 1H), 5.76-5.64 (m, 2H),5.35 (s, 2H), 5.17-5.11 (m, 4H), 4.98 (d, J=9.2, 1H), 4.87 (d, J=9.8 Hz,1H), 3.96 (m, J=10.4 Hz, 1H), 3.83-3.71 (m, 4H), 3.14 (dd, J=14.7, 4.3Hz, 1H), 3.0-2.87 (m, 1H), 2.55-2.35 (m, 9H), 2.11 (d, J=14.7 Hz, 1H),0.82 (s, 9H), 0.26 (s, 3H), 0.13 (s, 3H); MS (ESI) m/z 820.55 (M+H).

A solution of 2-5 (550 mg, 0.671 mmol, 1.0 equiv) in degassed CH₂Cl₂(2.5 mL, with 1 and 1.5 mL rinse) was added under nitrogen via syringeto a flame-dried flask containing N,N-dimethylbarbituric acid (324 mg,2.07 mmol, 3.0 equiv), and Tetrakis(triphenylphosphine)palladium(O)(56.9 mg, 0.0492 mmol, 0.07 equiv). The resulting solution was heated to35° C. for 4 h, then concentrated to remove the solvent. The resultingcrude mixture was purified via preparative reverse phase HPLC on aWaters Autopurification system using a Sunfire Prep C18 OBD column [5μm, 19×50 mm; flow rate, 20 mL/min; Solvent A: H₂O with 0.1% HCO₂H;Solvent B: CH₃CN with 0.1% HCO₂H; injection volume: 3×3.1 mL (CH₃CN);gradient: 80→100% B over 17 min; mass-directed fraction collection].Fractions with the desired MW, eluting at 6.1-10.1 min, were collectedand freeze-dried to give 352 mg of pure 2-6 (71%): ¹H NMR (400 MHz,CDCl₃) δ 16.10 (s, 1H), 7.51-7.43 (m, 4H), 7.9-7.29 (m, 6H), 6.61 (d,J=9.8 Hz, 1H), 5.35 (s, 2H), 4.87 (dd, J=22.6, 10.4 Hz, 2H), 3.96 (d,J=10.4 Hz, 1H), 3.91 (s, 2H), 3.12 (dd, J=15.3, 10.1 Hz, 1H), 3.04-2.92(m, 1H), 2.55-2.31 (m, 9H), 2.11 (d, J=14.7 Hz, 1H), 0.82 (s, 9H), 0.27(s, 3H), 0.12 (s, 3H); MS (ESI) m/z 740.44 (M+H).

To a solution of aniline 2-6 (30 mg, 0.041 mmol, 1 equiv) in THF (600μL) was added bromoacetylbromide (3.7 μL, 0.043 mmol, 1.05 equiv). After15 minutes, indazole (53 mg, 0.45 mmol, 10 equiv) was added. After 15 hthe reaction was heated to 80° C. After an additional 26 h, another 20mg indazole (0.17 mmol, 4 equiv) was added and the reaction heated at80° C. After 20 h, the solvent was removed in vacuo and the resultingcrude mixture dried under vacuum.

The above crude intermediate was transferred to a plastic vial indioxane (1.2 mL) and an aqueous solution of hydrogen fluoride (50%, 300μL) was added. After five hours, the reaction solution was diluted withan aqueous solution of K₂HPO₄ (3.6 g in 30 mL) and extracted with EtOAc(2×30 mL). The combined organic layers were dried (Na₂SO₄), filtered,and concentrated to yield the crude product.

Palladium on carbon (10%, 10 mg) was added to solution of the abovecrude intermediate in dioxane:methanol (1:1, 1 mL). The flask was fittedwith a septum and evacuated and back-filled three times with hydrogengas. Hydrogen gas was bubbled through the reaction solution for twominutes, then the reaction was stirred under an atmosphere (balloon) ofhydrogen gas for 1.5 h. The reaction mixture was filtered through celiteto remove the palladium catalyst and concentrated under reducedpressure. Preparative reverse phase HPLC purification of the resultingoil was performed on a Waters Autopurification system using a Polymerx10μ RP-γ 100 R column [30×21.20 mm, 10 micron, solvent A: 0.05N HCl inwater, solvent B: CH₃CN; injection volume: 4.8 mL (0.05N HCl in water);gradient elution with 10→60% B over 15 min, then held at 100% for 5 min;mass-directed fraction collection]. Fractions with the desired MW,eluting at 14-14.65 min, were collected and freeze-dried to yield 3.6 mgof compound 59 (15%): ¹H NMR (400 MHz, CD₃OD) δ 8.71 (s, 1H), 8.19 (d,J=11.0 Hz, 1H), 7.92-7.90 (m, 1H), 7.72-7.57 (m, 2H), 7/35-7.29 (m, 1H),5.65 (s, 2H), 4.08 (s, 1H), 3.16-2.92 (m, 9H), 2.31-2.18 (m, 2H),1.67-1.60 (m, 1H); MS (ESI) m/z 606.41 (M+H).

To a solution of aniline 2-6 (22 mg, 0.030 mmol, 1 equiv) in THF (500μL) was added bromoacetylbromide (2.7 μL, 0.031 mmol, 1.05 equiv). After30 minutes, pyrazole (36 mg, 0.53 mmol, 18 equiv) was added. After 20min the reaction was heated to 80° C. for 1.5 h, cooled to roomtemperature for 15 h, then heated at 80° C. for 4.5 h. The solvent wasremoved in vacuo and the resulting crude mixture dried under vacuum.

The above crude intermediate was transferred to a plastic vial inacetonitrile (1.0 mL) and an aqueous solution of hydrogen fluoride (50%,200 μL) was added. After 20 h, the reaction solution was diluted with anaqueous solution of K₂HPO₄ (2.4 g in 20 mL) and extracted with EtOAc(2×25 mL). The combined organic layers were dried (Na₂SO₄), filtered,and concentrated to yield the crude product.

Palladium on carbon (10%, 10 mg) was added to solution of the abovecrude intermediate in dioxane:methanol (1:1, 1 mL). The flask was fittedwith a septum and evacuated and back-filled three times with hydrogengas. Hydrogen gas was bubbled through the reaction solution for twominutes and the reaction was stirred under an atmosphere (balloon) ofhydrogen gas for 1.5 h. The reaction mixture was filtered through celiteto remove the palladium catalyst and concentrated under reducedpressure. Preparative reverse phase HPLC purification of the resultingoil was performed on a Waters Autopurification system using a Polymerx10μ RP-γ 100 R column [30×21.20 mm, 10 micron, solvent A: 0.05N HCl inwater, solvent B: CH₃CN; injection volume: 3.0 mL (10% CH₃CN in 0.05NHCl in water); gradient elution with 10→60% B over 10 min, then held at100% for 5 min; mass-directed fraction collection]. Fractions with thedesired MW, eluting at 8.8-10.2 min were collected and freeze-dried. Theresulting yellow solid was purified a second time via preparativereverse phase HPLC purification using the above procedure with agradient over 20 minutes, with the fractions with the desired MW elutingat 10.7-12.4 min were collected and freeze-dried to give 8.2 mg of pure60 (50%): ¹H NMR (400 MHz, CD₃OD) δ 8.19 (d, J=11.0 Hz, 1H), 8.05-7.92(m, 2H), 6.62-6.57 (m, 1H), 5.33 (d, J=4.9 Hz, 2H), 4.08 (s, 1H),3.16-2.90 (m, 9H), 2.31-2.17 (m, 2H), 1.69-1.55 (m, 1H); MS (ESI) m/z556.42 (M+H).

To a solution of aniline 2-6 (23 mg, 0.032 mmol, 1 equiv) in THF (500μL) was added bromoacetylbromide (2.9 μL, 0.034 mmol, 1.05 equiv). After30 minutes, imidazole (32 mg, 0.47 mmol, 15 equiv) was added and thesolution was heated to 80° C. After 2 h, the solution was cooled and thesolvent was removed in vacuo.

The above crude intermediate was transferred to a plastic vial indioxane (1.2 mL) and an aqueous solution of hydrogen fluoride (50%, 200μL) was added. After 1.5 h, the reaction solution was diluted with anaqueous solution of K₂HPO₄ (2.4 g in 30 mL) and extracted with EtOAc(2×25 mL). The combined organic layers were dried (Na₂SO₄), filtered,and concentrated to yield the crude product.

Palladium on carbon (10%, 8 mg) was added to solution of the above crudeintermediate in dioxane:methanol (1:1, 1 mL). The flask was fitted witha septum and evacuated and back-filled three times with hydrogen gas.Hydrogen gas was bubbled through the reaction solution and the reactionwas stirred under an atmosphere (balloon) of hydrogen gas for 1.5 h.More palladium catalyst was added and the evacuation and backfillingwith hydrogen was performed twice more at 1.5 h and 5 h. The reactionmixture was filtered through celite to remove the palladium catalyst andconcentrated under reduced pressure. Preparative reverse phase HPLCpurification of the resulting oil was performed on a WatersAutopurification system using a Polymerx 10μ RP-γ 100 R column [30×21.20mm, 10 micron, solvent A: 0.05N HCl in water, solvent B: CH₃CN;injection volume: 2.8 mL (0.05N HCl in water); gradient elution with10→60% B over 15 min, then held at 100% for 5 min; mass-directedfraction collection]. Fractions with the desired MW, eluting at 7.0-7.8min were collected and freeze-dried to give 4.1 mg of pure 61 (23%): ¹HNMR (400 MHz, CD₃OD) δ 9.02 (s, 1H), 8.17 (d, J=11.0 Hz, 1 H), 7.67 (s,1H), 7.61 (s, 1H), 5.34 (s, 2H), 4.09 (s, 1H), 3.18-2.90 (m, 9H),2.34-21.7 (m, 2H), 1.71-1.56 (m, 1H); MS (ESI) m/z 556.45 (M+H).

To a solution of aniline 2-6 (20.2 mg, 0.027 mmol, 1 equiv) in THF (500μL) was added bromoacetylbromide (2.5 μL, 0.029 mmol, 1.05 equiv). After30 minutes, 1H-1,2,3-triazole (31 μL, 0.54 mmol, 20 equiv) was added andthe solution was heated to 80° C. After 17 h, an additional 31 μL (20equiv) of 1H-1,2,3-triazole was added and the solution was heated for 22h. The solution was cooled and the solvent was removed in vacuo.

The above crude intermediate was transferred to a plastic vial indioxane (1.0 mL) and an aqueous solution of hydrogen fluoride (50%, 200μL) was added. After 17 h, the reaction solution was diluted with anaqueous solution of K₂HPO₄ (2.4 g in 20 mL) and extracted with EtOAc(2×25 mL). The combined organic layers were dried (Na₂SO₄), filtered,and concentrated to yield the crude product.

Palladium on carbon (10%, 7 mg) was added to solution of the above crudeintermediate in dioxane:methanol (1:1, 1 mL). The flask was fitted witha septum and evacuated and back-filled three times with hydrogen gas.Hydrogen gas was bubbled through the reaction solution and the reactionwas stirred under an atmosphere (balloon) of hydrogen gas for 1.5 h. Thereaction mixture was filtered through celite to remove the palladiumcatalyst and concentrated under reduced pressure. Preparative reversephase HPLC purification of the resulting oil was performed on a WatersAutopurification system using a Polymerx 10μ RP-γ 100 R column [30×21.20mm, 10 micron, solvent A: 0.05N HCl in water, solvent B: CH₃CN;injection volume: 2.5 mL (0.05N HCl in water); gradient elution with10→60% B over 15 min, then held at 100% for 5 min; mass-directedfraction collection]. Fractions with the desired MW, eluting at9.25-10.5 min were collected and freeze-dried. Purification wasperformed a second time as above and the fractions with the desired MW,eluting at 9.75-10.25 min were collected and freeze-dried to give 1.5 mgof pure 62 (10%): ¹H NMR (400 MHz, CD₃OD) δ 8.24 (s, 1H), 8.17 (d,J=11.0 Hz, 1H), 8.00 (s, 1H), 5.57 (s, 2H), 4.09 (s, 1H), 3.16-2.92 (m,9H), 2.34-2.16 (m, 2H), 1.71-1.67 (m, 1H); MS (ESI) m/z 557.44 (M+H).

To a solution of aniline 2-6 (16.7 mg, 0.023 mmol, 1 equiv) in THF (500μL) was added bromoacetylbromide (2.0 μL, 0.024 mmol, 1.05 equiv). After20 minutes, a tetrazole solution (0.45M in CH₃CN, 500 μL, 0.23 mmol, 10equiv) was added and the solution was heated to 80° C. After 4 h,potassium carbonate (35 mg, 0.25 mmol, 11 equiv) was added and thereaction heated for 35 minutes. The solution was cooled and filteredthrough celite, and the solvent was removed in vacuo.

The above crude intermediate was transferred to a plastic vial indioxane (1.0 mL) and an aqueous solution of hydrogen fluoride (50%, 200μL) was added. After 18 h, the reaction solution was diluted with anaqueous solution of K₂HPO₄ (2.4 g in 20 mL) and extracted with EtOAc(2×25 mL). The combined organic layers were dried (Na₂SO₄), filtered,and concentrated to yield the crude product.

Palladium on carbon (10%, 7 mg) was added to solution of the above crudeintermediate in dioxane:methanol (1:1, 1 mL). The flask was fitted witha septum and evacuated and back-filled three times with hydrogen gas.Hydrogen gas was bubbled through the reaction solution and the reactionwas stirred under an atmosphere (balloon) of hydrogen gas for 1 h. Thereaction mixture was filtered through celite to remove the palladiumcatalyst and concentrated under reduced pressure. Preparative reversephase HPLC purification of the resulting oil was performed on a WatersAutopurification system using a Polymerx 10μ RP-γ 100 R column [30×21.20mm, 10 micron, solvent A: 0.05N HCl in water, solvent B: CH₃CN;injection volume: 2.5 mL (10% CH₃CN in 0.05N HCl in water); gradientelution with 10→60% B over 15 min, then held at 100% for 5 min;mass-directed fraction collection]. Fractions with the desired MW,eluting at 11.2-12.1 min were collected and freeze-dried. Purificationwas performed a second time as above with the gradient extended over 20min. Fractions with the desired MW, eluting at 13.7-14.5 min werecollected and freeze-dried to give 1.6 mg of pure 63 (13%): ¹H NMR (400MHz, CD₃OD) δ 8.78 (s, 1H), 8.14 (d, J=11.0 Hz, 1H), 5.78 (s, 2H), 4.07(s, 1H), 3.17-2.81 (m, 9H), 2.36-2.16 (m, 2H), 1.70-1.52 (m, 1H); MS(ESI) m/z 558.43 (M+H).

Example 3. Synthesis of Compounds of Structural Formula (A), wherein Xis hydrogen and Y is —NH—C(O)-heterocyclyl, —NH—C(O)-heteroaryl—NH—C(O)—[C(R^(D))(R^(E))]₀₋₁—N(R^(A))(R^(B)), —NH—C(O)-carbocyclyl,—NH—C(O)-aryl, —NH—SO₂—(C₁-C₆)alkyl, —NH—SO₂-carbocyclyl, —NH—SO₂-aryl,—NH—SO₂-heterocyclyl or —NH—SO₂-heteroaryl

In Scheme 3, R′ represents heterocyclyl, heteroaryl, carbocyclyl, aryl,(C₁-C₆)alkyl, or -[(C(R^(D))(R^(E))]₀₋₁—N(R^(A))(R^(B)); and R^(2′)represents hydrogen, (C₁-C₆)alkyl, —(C₀-C₅)alkylene-carbocyclyl,—(C₀-C₅)alkylene-aryl, —(C₀-C₅)alkylene-heteroaryl, or—(C₀-C₅)alkylene-heterocyclyl. For certain compounds made by Scheme 3and described below, R^(Z) is hydrogen and R^(X) and R^(Y) are takentogether with the carbon and nitrogen atoms to which they are bound toform an optionally substituted 4-7 membered saturated heterocyclyl. Itwill be readily apparent to those of skill in the art, however, thatthis Scheme 3 will also be useful to synthesize compounds where R^(X),R^(Y) and R^(Z) are R^(B), R^(D) and R^(E), respectively, as defined instructural formula (A). The following compounds were prepared accordingto Scheme 3.

To a solution of aniline 9 (17.0 mg, 0.038 mmol, 1 equiv) in DMF (200μL) was added N-Benzyloxycarbonyl-L-proline acid chloride (1.0 M intoluene, 57 μL, 1.5 equiv). After 50 min the reaction mixture wasdiluted to 3 mL with 0.05 N HCl in H₂O and filtered to remove anysolids. Preparative reverse phase HPLC purification of the resultingsolution was performed on a Waters Autopurification system using aPolymerx 10μ RP-γ 100 R column [30×21.20 mm, 10 micron, solvent A: 0.05NHCl in water, solvent B: CH₃CN; injection volume: 3.5 mL (0.05N HCl inwater); gradient elution with 10→20% B over 25 min, then held at 100%for 5 min; mass-directed fraction collection]. Fractions with thedesired MW, eluting at 27.1-28.4 min, were collected and freeze-dried.

Palladium on carbon (10%, 10 mg) was added to a solution of the aboveintermediate in dioxane:MeOH (1:3, 2.3 mL). The flask was fitted with aseptum and evacuated and back-filled three times with hydrogen gas. Thereaction solution was stirred under an atmosphere (balloon) of hydrogengas for 1.7 h, then was filtered through celite to remove the palladiumcatalyst and concentrated under reduced pressure. Half of the resultingresidue was purified via preparative reverse phase HPLC purification ona Waters Autopurification system using a Polymerx 10μ RP-γ 100 R column[30×21.20 mm, 10 micron, solvent A: 0.05N HCl in water, solvent B:CH₃CN; injection volume: 1.8 mL (0.05 N HCl in water); gradient elutionwith 0→35% B over 10 min, then held at 100% for 5 min; mass-directedfraction collection]. Fractions with the desired MW, eluting at 7.8-8.5min, were collected and freeze-dried to yield 1.9 mg of compound 64(30%): ¹H NMR (400 MHz, CD₃OD) δ 8.16 (d, J=11.0 Hz, 1H), 4.59-4.56 (m,1H), 4.10 (s, 1H), 3.48-3.33 (m, 2H), 3.18-2.95 (m, 9H), 2.59-2.50 (m,1H), 2.34-2.05 (m, 5H), 1.70-1.60 (m, 1H); MS (ESI) m/z 545.38 (M+H).

To a solution of aniline 9 (15.7 mg, 0.035 mmol, 1 equiv) in DMF (200μL) was added N-Benzyloxycarbonyl-D-proline acid chloride (1.0 M intoluene, 53 μL, 1.5 equiv). After 50 min, the reaction was complete. Thereaction mixture was diluted to 3 mL with 0.05 N HCl in H₂O and filteredto remove any solids. Preparative reverse phase HPLC purification of theresulting solution was performed on a Waters Autopurification systemusing a Polymerx 10μ RP-γ 100 R column [30×21.20 mm, 10 micron, solventA: 0.05 N HCl in water, solvent B: CH₃CN; injection volume: 3.5 mL (0.05N HCl in water); gradient elution with 15→80% B over 10 min, then heldat 100% for 5 min; mass-directed fraction collection]. Fractions withthe desired MW, eluting at 6.95-8.10 min, were collected andfreeze-dried.

Palladium on carbon (10%, 15 mg) was added to a solution of the aboveintermediate in dioxane:MeOH (1:3, 2.3 mL). The flask was fitted with aseptum and evacuated and back-filled three times with hydrogen gas, andthe reaction was stirred under an atmosphere (balloon) of hydrogen gasfor 1.5 h. The reaction mixture was filtered through celite to removethe palladium catalyst and concentrated under reduced pressure. Half ofthe resulting residue was purified via preparative reverse phase HPLCpurification on a Waters Autopurification system using a Polymerx 10μRP-γ 100 R column [30×21.20 mm, 10 micron, solvent A: 0.05 N HCl inwater, solvent B: CH₃CN; injection volume: 1.8 mL (0.05 N HCl in water);gradient elution with 0→35% B over 10 min, then held at 100% for 5 min;mass-directed fraction collection]. Fractions with the desired MW,eluting at 8.35-8.85 min, were collected and freeze-dried to yield 0.93mg of compound 65 (24%): ¹H NMR (400 MHz, CD₃OD) δ 8.17 (d, J=11.0 Hz,1H), 4.59-4.53 (m, 1H), 4.09 (s, 1H), 3.48-3.37 (m, 2H), 3.18-2.90 (m,9H), 2.59-2.50 (m, 1H), 2.34-2.05 (m, 5H), 1.70-1.59 (m, 1H); MS (ESI)m/z 545.37 (M+H).

The second half of crude 64 (0.012 mmol, 1.0 equiv) was dissolved in DMF(500 μL), and formaldehyde (37% aqueous solution, 5.3 μL, 0.072 mmol, 6equiv), triethylamine (5.0 μL, 0.036 mmol, 3 equiv), and sodiumtriacetoxyborohydride (8.4 mg, 0.039 mmol, 3.2 equiv) were addedsequentially. After 2 h, the reaction mixture was diluted to 1.8 mL with0.05N HCl in H₂O and purified via preparative reverse phase HPLC on aWaters Autopurification system using a Polymerx 10μ RP-γ 100 R column[30×21.20 mm, 10 micron, solvent A: 0.05 N HCl in water, solvent B:CH₃CN; injection volume: 1.8 mL (0.05 N HCl in water); gradient: 0→30% Bover 10 min; mass-directed fraction collection]. Fractions with thedesired MW, eluting at 8.6-9.35 min, were collected and freeze-dried toprovide a mixture of the desired compound and an over-formylatedproduct. The resulting compound mixture was dissolved in 4 N aqueous HClsolution (1.5 mL) and stirred for 50 h, then freeze-dried to provide 1.0mg of the desired compound 66 (15%): ¹H NMR (400 MHz, CD₃OD) δ 8.17 (d,J=10.4 Hz, 1H), 4.36 (t, J=8.6 Hz, 1H), 4.08 (s, 1H), 3.82-3.73 (m, 1H),3.20-2.90 (m, 12H), 2.73-2.68 (m, 1H), 2.35-2.10 (m, 5H), 1.70-1.60 (m,1H); MS (ESI) m/z 559.38 (M+H).

The second half of crude 65 (0.007 mmol, 1.0 equiv) was dissolved in DMF(500 μL), and formaldehyde (37% aqueous solution, 3.1 μL, 0.042 mmol, 6equiv) and TEA (3.0 μL, 0.021 mmol, 3 equiv), and sodiumtriacetoxyborohydride (4 mg, 0.026 mmol, 2.6 equiv) were addedsequentially. After 2.2 h, the reaction mixture was diluted to 1.8 mLwith 0.05N HCl in H₂O and purified via preparative reverse phase HPLC ona Waters Autopurification system using a using a Polymerx 10μ RP-γ 100 Rcolumn [30×21.20 mm, 10 micron, solvent A: 0.05 N HCl in water, solventB: CH₃CN; injection volume: 2.0 mL (0.05 N HCl in water); gradient:0→30% B over 10 min; mass-directed fraction collection]. Fractions withthe desired MW, eluting at 8.9-9.6 min, were collected and freeze-driedto provide a mixture of the desired compound and an over-formylatedproduct. The resulting compound mixture was dissolved in 6 N aqueous HClsolution and stirred for 50 h, then freeze-dried to provide 1.5 mg ofthe desired compound 67 (38%): ¹H NMR (400 MHz, CD₃OD) δ 8.17 (d, J=10.4Hz, 1H), 4.45-4.34 (m, 1H), 4.08 (s, 1H), 3.84-3.74 (m, 1H), 3.20-2.90(m, 12H), 2.79-2.65 (m, 1H), 2.33-2.05 (m, 5H), 1.70-1.58 (m, 1H); MS(ESI) m/z 559.40 (M+H).

To a solution of (S)-(−)-1-Cbz-piperidinecarboxylic acid (34.2 mg, 0.13mmol, 3 equiv), and(2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluoro-phosphate) (50.0 mg, 0.13 mol, 3 equiv) in DMF (200 μL) wasadded triethylamine (18 μL, 0.13 mmol, 3 equiv). After 30 min, aniline 9(17.5 mg, 0.039 mmol, 1 equiv) was added. After 16 h, the reactionmixture was diluted to 3 mL with 0.05 N HCl in H₂O and purified viapreparative reverse phase HPLC on a Waters Autopurification system usinga Polymerx 10μ RP-γ 100 R column [30×21.20 mm, 10 micron, solvent A:0.05 N HCl in water, solvent B: CH₃CN; injection volume: 3.5 mL (0.05 NHCl in water); gradient elution with 15→70% B over 10 min, then held at100% for 5 min; mass-directed fraction collection]. Fractions with thedesired MW, eluting at 9.07-10.0 min, were collected and freeze-dried.Palladium on carbon (10%, 4 mg) was added to a solution of this foam indioxane:MeOH (1:3, 1.2 mL). The flask was fitted with a septum andevacuated and back-filled three times with hydrogen gas. The reactionmixture was stirred under an atmosphere (balloon) of hydrogen gas for1.5 h, then was filtered through celite to remove the palladium catalystand concentrated under reduced pressure. Preparative reverse phase HPLCpurification of the resulting oil was performed on a WatersAutopurification system using a Polymerx 10μ RP-γ 100 R column [30×21.20mm, 10 micron, solvent A: 0.05 N HCl in water, solvent B: CH₃CN;injection volume: 2.0 mL (0.05 N HCl in water); gradient elution with0→35% B over 10 min, then held at 100% for 5 min; mass-directed fractioncollection]. Fractions with the desired MW, eluting at 8.15-8.58 min,were collected and freeze-dried to yield 0.75 mg of compound 68 (4%): ¹HNMR (400 MHz, CD₃OD) δ 8.15 (d, J=11.0 Hz, 1H), 4.12-4.06 (m, 2H),3.48-3.40 (m, 2H), 3.20-2.90 (m, 9H), 2.36-2.18 (m, 3H), 2.02-1.90 (m,2H), 1.82-1.60 (m, 4H); MS (ESI) m/z 559.37 (M+H).

To a solution of (R)-(+)-1-Cbz-piperidinecarboxylic acid (35.0 mg, 0.13mmol, 3 equiv), and(2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluoro-phosphate) (50.0 mg, 0.13 mol, 3 equiv) in DMF (200 μL) wasadded TEA (18 μL, 0.13 mmol, 3 equiv). After 30 min, aniline 9 (16.6 mg,0.037 mmol, 1 equiv) was added. After 16 h, the reaction mixture wasdiluted to 3 mL with 0.05 N HCl in H₂O and purified via preparativereverse phase HPLC on a Waters Autopurification system using a Polymerx10μ RP-γ 100 R column [30×21.20 mm, 10 micron, solvent A: 0.05 N HCl inwater, solvent B: CH₃CN; injection volume: 3.5 mL (0.05 N HCl in water);gradient elution with 10→50% B over 10 min, then held at 100% for 5 min;mass-directed fraction collection]. Fractions with the desired MW,eluting at 12.1-12.9 min, were collected and freeze-dried. Palladium oncarbon (10%, 5 mg) was added to a solution of this foam in dioxane:MeOH(1:3, 800 μL). The flask was fitted with a septum and evacuated andback-filled three times with hydrogen gas. The reaction mixture wasstirred under an atmosphere (balloon) of hydrogen gas for 1.75 h, thenwas filtered through celite to remove the palladium catalyst andconcentrated under reduced pressure. Preparative reverse phase HPLCpurification of the resulting oil was performed on a WatersAutopurification system using a Polymerx 10μ RP-γ 100 R column [30×21.20mm, 10 micron, solvent A: 0.05 N HCl in water, solvent B: CH₃CN;injection volume: 2.0 mL (0.05 N HCl in water); gradient elution with0→35% B over 10 min, then held at 100% for 5 min; mass-directed fractioncollection]. Fractions with the desired MW, eluting at 8.75-9.16 min,were collected and freeze-dried to yield 0.55 mg of compound 69 (3%): ¹HNMR (400 MHz, CD₃OD) δ 8.16 (d, J=11.0 Hz, 1H), 4.13-4.06 (m, 2H),3.50-3.43 (m, 2H), 3.20-2.90 (m, 9H), 2.38-2.18 (m, 3H), 2.04-1.88 (m,2H), 1.83-1.60 (m, 4H); MS (ESI) m/z 559.38 (M+H).

To a solution of compound 68 (0.0138 mmol, 1 equiv) in DMF (750 μL),were added formaldehyde (37% aqueous solution, 6.2 μL, 0.083 mmol, 6equiv), TEA (5.8 μL, 0.041 mmol, 3 equiv), and sodiumtriacetoxyborohydride (11 mg, 0.051 mmol, 3.7 equiv) sequentially. After17 h, the reaction mixture was concentrated to remove amine and 6 Naqueous HCl (500 μL) was added. After 19 days, the reaction solution waspurified via preparative reverse phase HPLC on a Waters Autopurificationsystem using a Polymerx 10μ RP-γ 100 R column [30×21.20 mm, 10 micron,solvent A: 0.05 N HCl in water, solvent B: CH₃CN; injection volume: 2.5mL (0.05 N HCl in water); gradient: 15→50% B over 15 min; mass-directedfraction collection]. Fractions with the desired MW, eluting at 5.75-6.2min, were collected and freeze-dried to provide 2.4 mg of the desiredcompound 70 (31%): ¹H NMR (400 MHz, CD₃OD) δ 8.16 (d, J=11.0 Hz, 1H),4.08-4.04 (m, 1H), 3.59-3.53 (m, 1H), 3.20-3.10 (m, 5H), 3.06-2.96 (m,5H), 2.90m (s, 3H), 2.36-2.25 (m, 2H), 2.11-2.05 (m, 1H), 2.02-1.94 (m,2H), 1.90-1.74 (m, 2H), 1.71-1.58 (m, 2H); MS (ESI) m/z 573.33 (M+H).

Compound 9 (20 mg, 0.045 mmol, 1.0 equiv) in THF was added Na₂CO₃ (9.5mg, 0.089 mmol, 2.0 equiv), (4R)-4-fluoro-1-methyl-L-proline (9.8 mg,0.067 mmol, 1.5 equiv) and HATU (34.6 mg, 0.047 mmol, 2.0 equiv). Thereaction mixture was stirred at room temperature for 20 hour. LC-MSanalysis indicated the starting material was consumed completely.HCl/MeOH (1 mL, 4 N) was added to the mixture at 0° C. and stirred for 2min. The mixture was concentrated under vacuum, the residue was purifiedby reverse phase HPLC to afford product 71 (6.1 mg): ¹H NMR (400 MHz,CD₃OD) δ 8.18 (d, J=10.8 Hz, 1H), 5.51 (d, J=51.6 Hz, 1H), 4.76-4.72 (m,1H), 4.22-4.16 (m, 1H), 4.10 (s, 1H), 3.74-3.63 (m, 1H), 3.21-2.97 (m,14H), 2.35-2.21 (m, 2H), 1.69-1.60 (m, 1H); MS (ESI) m/z 577.1 (M+H).

Compounds 72 and 73 were prepared similarly to compound 71 using thecorresponding amino acids.

Prepared similarly to compound 71: ¹H NMR (400 MHz, CD₃OD) δ 8.16 (d,J=10.8 Hz, 1H), 5.48 (d, J=51.2 Hz, 1H), 4.60-4.56 (m, 1H), 4.11 (s,1H), 4.05-3.98 (m, 1H), 3.67-3.54 (m, 1H), 3.24-2.96 (m, 13H), 2.55-2.44(m, 1H), 2.34-2.22 (m, 2H), 1.70-1.66 (m, 1H); MS (ESI) m/z 577.1 (M+H).

Prepared similarly to compound 71: ¹H NMR (400 MHz, CD₃OD) δ 8.18 (d,J=10.8 Hz, 1H), 4.76-4.71 (m, 1H), 4.17-4.12 (m, 1H), 4.09 (s, 1H),3.96-3.86 (m, 1H), 3.67-3.53 (m, 1H), 3.55-3.53 (m, 1H), 3.25-2.73 (m,12H), 2.33-2.19 (m, 2H), 1.68-1.59 (m, 1H); MS (ESI) m/z 595.3 (M+H).

1-(Bocamino)cyclopropanecarboxylic acid (67.4 mg, 0.335 mmol),O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (127 mg, 0.335 mmol), and triethylamine (0.078 mL,0.56 mmol) were stirred in DMF (1 mL) for 30 minutes. Compound 9 (50 mg,0.112 mmol) was added. After stirring overnight, the reaction mixturewas purified directly by preparative reverse phase HPLC purification ona Waters Autopurification system using a Polymerx 10μ RP-γ 100 R column[30×21.20 mm, 10 micron, solvent A: 0.05N HCl, solvent B: CH₃CN,gradient elution with 0→50% B over 10 min; mass-directed fractioncollection]. Fractions with the desired MW were collected andfreeze-dried. The material was repurified by preparative reverse phaseHPLC purification on a Waters Autopurification system using a PolymerxRP-γ 100 R column [30×21.20 mm, 10 micron, solvent A: 0.05N HCl, solventB: CH₃CN, gradient elution with 0→100% B over 15 min; mass-directedfraction collection]. Fractions with the desired MW were collected andfreeze-dried. This gave 42 mg of compound 3-1-1 (59%, ˜80% pure) whichwas used without further purification: MS (ESI) m/z 631.41 (M+H).

Compound 3-1-1 (42 mg, 0.067 mmol, ˜80% pure) was stirred in 4M HCl in1,4-dioxane (5 mL) overnight. The reaction mixture was concentratedunder reduced pressure and was purified by preparative reverse phaseHPLC purification on a Waters Autopurification system using a Polymerx10μ RP-γ 100 R column [30×21.20 mm, 10 micron, solvent A: 0.05N HCl,solvent B: CH₃CN, gradient elution with 0→50% B over 10 min;mass-directed fraction collection]. Fractions with the desired MW werecollected and freeze-dried. The material was dissolved in MeOH (1 mL),and the solution was added dropwise to vigorously stirring diethyl ether(200 mL). The resulting solid was collected by filtration on a pad ofCelite. This was washed with diethyl ether (3 times), and the solid wasdissolved in MeOH and concentrated under reduced pressure. The materialwas freeze-dried, yielding 25.8 mg of compound 74: ¹H NMR (400 MHz,CD₃OD with 1 drop DCl) δ 8.00 (d, J=7.0 Hz, 1H), 4.05 (s, 1H), 3.20-2.85(m, 9H), 2.36-2.06 (m, 2H), 1.70-1.52 (m, 3H), 1.35-1.22 (m, 2H); MS(ESI) m/z 531.33 (M+H).

To a dichloromethane (5 mL) suspension of compound 9 (0.260 g, 0.50mmol, 1.0 equiv) at rt was added triethylamine (0.139 mL, 1.00 mmol, 2.0equiv). The reaction was stirred at rt until form a clear solution.Methylisocyanate (89.4 μL, 1.50 mmol, 3.0 equiv) was added to thereaction mixture dropwise. The reaction was allowed to stir at 25° C.for 1 h. Additional methylisocyanate (45 μL, 0.75 mmol, 1.5 equiv) wasadded and stirred overnight. LCMS indicate there are still startingmaterial present. The solvent was removed under vacuum to give the crude75. The crude product was purified by HPLC on a Polymerx 10μ RP-γ 100 Rcolumn [30×21.20 mm, 10 micron, solvent A: 0.05 N HCl, solvent B: CH₃CN,sample in 2.0 mL (0.05 N HCl), gradient elution with 15→65% B over 15min, mass-directed fraction collection] to yield the desired product 75as a yellow solid (80 mg, 31.7%): ¹H NMR (400 MHz, CD₃OD) δ 8.12 (d,J=11.4 Hz, 1H), 4.07 (s, 1H), 3.04 (s, 3H), 2.96 (s, 3H), 3.13-2.93 (m,3H), 2.77 (s, 3H), 2.27-2.15 (m, 2H), 1.69-1.57 (m, 1H); MS (ESI) m/z505.41 (M+H).

Compound 9 (20 mg, 0.045 mmol, 1.0 equiv) in THF was added Na₂CO₃ (9.5mg, 0.089 mmol, 2.0 equiv) and 0.1 mL benzoyl chloride solution (54 μLin 1 mL THF, 0.047 mmol, 1.05 equiv). The reaction mixture was stirredat room temperature for 1 hour. LC-MS analysis indicated the startingmaterial was consumed completely. HCl/MeOH (1 mL, 4 N) was added to themixture at 0° C. and stirred for 2 min. The mixture was concentratedunder vacuum, the residue was purified by reverse phase HPLC to affordproduct 76 (5.5 mg): ¹H NMR (400 MHz, CD₃OD) δ 8.23 (d, J=10.8 Hz, 1H),7.97 (d, J=7.6 Hz, 2H), 7.66-7.54 (m, 3H), 4.11 (s, 1H), 3.21-2.90 (m,9H), 2.37-2.24 (m, 2H), 1.72-1.66 (m, 1H); MS (ESI) m/z 552.1 (M+H).

Compounds 77-83 were prepared similarly to compound 76 using thecorresponding acid chlorides.

¹H NMR (400 MHz, CD₃OD) δ 8.25 (s, 1H), 8.21 (d, J=8.0 Hz, 1H), 8.14 (d,J=10.4 Hz, 1H), 7.92 (d, J=8.0 Hz, 1H), 7.76 (t, J=8.0 Hz, 1H), 4.08 (s,1H), 3.21-2.89 (m, 9H), 2.35-2.22 (m, 2H), 1.71-1.61 (m, 1H); MS (ESI)m/z 620.1 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.10 (d, J=10.8 Hz, 1H), 7.41-7.33 (m, 3H),7.09-7.07 (m, 1H), 4.00 (s, 1H), 3.78 (s, 3H), 3.12-2.86 (m, 9H),2.23-2.13 (m, 2H), 1.60-1.50 (m, 1H); MS (ESI) m/z 582.1 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.12 (d, J=10.8 Hz, 1H), 7.89 (d, J=3.2 Hz,1H), 7.78 (d, J=4.8 Hz, 1H), 7.22 (t, J=8.8 Hz, 1H), 4.10 (s, 1H),3.20-2.98 (m, 9H), 2.36-2.20 (m, 2H), 1.68-1.61 (m, 1H); MS (ESI) m/z558.1 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 9.34 (s, 1H), 9.04-9.00 (m, 2H), 8.20-8.15 (m,2H), 4.07 (s, 1H), 3.27-2.94 (m, 9H), 2.34-2.18 (m, 2H), 1.68-1.59 (m,1H); MS (ESI) m/z 553.1 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.13-8.06 (m, 2H), 7.98 (d, J=7.6 Hz, 1H),7.77 (d, J=7.2 Hz, 1H), 7.67 (t, J=8.0 Hz, 1H), 4.01 (s, 1H), 3.26 (s,6H), 3.14-2.83 (m, 9H), 2.27-2.13 (m, 2H), 1.64-1.52 (m, 1H); MS (ESI)m/z 595.1 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.08 (d, J=10.8 Hz, 1H), 7.98 (d, J=8.4 Hz,2H), 7.49 (d, J=8.4 Hz, 2H), 4.02 (s, 1H), 3.19 (s, 6H), 3.12-2.88 (m,9H), 2.24-2.13 (m, 2H), 1.60-1.51 (m, 1H); MS (ESI) m/z 595.1 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 8.19-8.14 (m, 2H), 8.05 (d, J=8.4 Hz, 1H),7.91-7.89 (m, 1H), 7.76-7.74 (m, 1H), 4.12 (s, 1H), 3.32 (s, 6H),3.21-2.96 (m, 9H), 2.41-1.98 (m, 2H), 1.72-1.59 (m, 1H); MS (ESI) m/z595.0 (M+H).

Compound 9 (20 mg, 0.045 mmol, 1.0 equiv) in THF was added DIEA (11.5mg, 0.089 mmol, 2.0 equiv) and 2-thiophenesulfonyl chloride (12.2 mg,0.067 mmol, 1.5 equiv). The reaction mixture was stirred at roomtemperature for 20 hour. LC-MS analysis indicated the starting materialwas consumed completely. HCl/MeOH (1 mL, 4 N) was added to the mixtureat 0° C. and stirred for 2 min. The mixture was concentrated undervacuum, the residue was purified by reverse phase HPLC to affordcompound 84 (2.0 mg): ¹H NMR (400 MHz, CD₃OD) δ 7.75 (dd, J=5.2, 1.2 Hz,1H), 7.59 (d, J=2.8 Hz, 1H), 7.52 (d, J=10.4 Hz, 1H), 7.09 (t, J=4.4 Hz,1H), 4.07 (s, 1H), 3.11-2.92 (m, 9H), 2.30-2.18 (m, 2H), 1.68-1.58 (m,1H); MS (ESI) m/z 593.9 (M+H).

Compounds 85-87 were prepared similarly to compound 84 using thecorresponding sulfonyl chlorides.

¹H NMR (400 MHz, CD₃OD) δ 7.44 (d, J=10.0 Hz, 1H), 4.10 (s, 1H),3.21-2.90 (m, 12H), 2.34-2.22 (m, 2H), 1.67-1.61 (m, 1H); MS (ESI) m/z526.1 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 7.82 (d, J=7.6 Hz, 2H), 7.58-7.46 (m, 4H),4.07 (s, 1H), 3.10-2.92 (m, 9H), 2.35-2.25 (m, 2H), 1.65-1.55 (m, 1H);MS (ESI) m/z 552.1 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 7.72 (t, J=5.6 Hz 1H), 7.62 (d, J=7.6 Hz, 1H),7.50 (d, J=8.4 Hz, 1H), 7.41-7.38 (m, 2H), 3.97 (s, 1H), 3.03-2.82 (m,9H), 2.19-2.06 (m, 2H), 1.53-1.50 (m, 1H); MS (ESI) m/z 622.1 (M+H).

Example 4. Synthesis of Compounds of Structural Formula (I), wherein Xis hydrogen and Y is —NH—C(O)-heterocycyl or —NH—C(O)-heteroaryl

In Scheme 4, R represents heteroaryl and R² is R^(A) as defined inStructural Formula (A). For certain compounds made by Scheme 4 anddescribed below, R^(Z) is hydrogen and R^(X) and R^(Y) are takentogether with the carbon and nitrogen atoms to which they arerespectively bound to form an optionally substituted 4-7 memberedsaturated heterocyclyl. It will be readily apparent to those of skill inthe art, however, that this Scheme 4 will also be useful to synthesizecompounds where R^(X), R^(Y) and R^(Z) are R^(B), R^(D) and R^(E),respectively, as defined in structural formula (A).

The following compounds were prepared according to Scheme 4.

To a suspension of 1-Fmoc-L-azetidine-2-carboxylic acid (135 mg, 0.42mmol, 2.9 equiv), and(2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluoro-phosphate) (164 mg, 0.43 mol, 3 equiv) in THF (1.5 mL) wasadded triethylamine (60 μL, 0.43 mmol, 3 equiv). After 30 min, aniline2-6 (106 mg, 0.14 mmol, 1 equiv) was added. After 18 h, the reactionmixture was concentrated under reduced pressure. Preparative reversephase HPLC of the resulting oil was performed on a WatersAutopurification system using a Sunfire Prep C18 OBD column [5 μm, 19×50mm; flow rate, 20 mL/min; Solvent A: H₂O with 0.1% HCO₂H; Solvent B:CH₃CN with 0.1% HCO₂H; injection volume: 3×2.0 mL (CH₃CN); gradient:80→100% B over 15 min; mass-directed fraction collection]. Fractionswith the desired MW, eluting at 10.35-12.0 min, were collected andfreeze-dried to provide 131 mg of a yellow powder.

To a solution of the above intermediate in CH₂Cl₂ (2 mL) was addedpiperidine (500 μL). After 30 min, the reaction solution was poured intoaqueous pH 7 phosphate buffer and extracted with EtOAc (3×20 mL). Thecombined organic layers were dried (Na₂SO₄), filtered, and concentratedunder reduced pressure. Purification of the resulting crude oil viaflash column chromatography on silica gel (Silicycle, 5 g, 0 to 5 to 10to 50% EtOAc in hexane gradient) provided 47.6 mg of the intermediate.

Half of the above intermediate (24 mg) was dissolved in acetonitrile (1mL), and an aqueous solution of HF (50%, 200 μL) was added. After 18.5h, the reaction solution was poured into an aqueous K₂HPO₄ solution (2.5g in 20 mL) and extracted with EtOAc (2×25 mL). The combined organiclayers were dried (Na₂SO₄), filtered, and concentrated under reducedpressure.

Palladium on carbon (10%, 12.5 mg) was added to a solution of the aboveintermediate in dioxane:MeOH (1:1, 1 mL). The flask was fitted with aseptum and evacuated and back-filled three times with hydrogen gas.Hydrogen gas was bubbled through the reaction solution for threeminutes, and the reaction mixture was stirred under an atmosphere(balloon) of hydrogen gas for 4.5 h. The reaction mixture was filteredthrough celite to remove the palladium catalyst and concentrated underreduced pressure. Preparative reverse phase HPLC purification of theresulting oil was performed on a Waters Autopurification system using aPolymerx 10μ RP-γ 100 R column [30×21.20 mm, 10 micron, solvent A: 0.05NHCl in water, solvent B: CH₃CN; injection volume: 3.0 mL (0.05N HCl inwater); gradient elution with 0→30% B over 10 min, then held at 100% for5 min; mass-directed fraction collection]. Fractions with the desiredMW, eluting at 9.8-11.25 min, were collected and freeze-dried. Theresulting impure powder was purified via preparative reverse phase HPLCas above with gradient elution with 15→50% B over 12 min, then held at100% for 3 min; mass-directed fraction collection. Fractions with thedesired MW, eluting at 6.5-8.0 min, were collected and freeze-dried toyield 2.0 mg of compound 88 (5%): ¹H NMR (400 MHz, CD₃OD) δ 8.25 (d,J=11.0 Hz, 1H), 5.29-5.24 (m, 1H), 4.20-4.11 (m, 1H), 4.09 (s, 1H),3.19-2.89 (m, 10H), 2.69-2.56 (m, 1H), 2.33-2.19 (m, 2H), 1.68-1.56 (m,1H); MS (ESI) m/z 531.30 (M+H).

N-methyl-L-azetidine-2-carboxylic acid

To a suspension of L-azetidine-2-carboxylic acid (290 mg, 2.87 mmol, 1equiv) in MeOH (3.6 mL), was added aqueous formaldehyde solution (37%,235 μL, 3.15 mmol, 1.1 equiv) and palladium on carbon (10%, 76 mg). Theflask was fitted with a septum and evacuated and back-filled three timeswith hydrogen gas. The reaction was stirred under an atmosphere(balloon) of hydrogen gas for 19 h, and was filtered through celite toremove the palladium catalyst. The resulting solution was concentratedunder reduced pressure, concentrated from toluene three times and driedunder vacuum to afford N-methyl-L-azetidine-2-carboxilic acid: ¹H NMR(400 MHz, CD₃OD) δ 4.50 (t, J=9.5 Hz, 1H), 3.96 (dt, J=4.3, 9.8 Hz, 1H),3.81 (q, J=9.8 Hz, 1H), 2.86 (s, 3H), 2.71-2.60 (m, 1H), 2.50-2.38 (m,1H).

To a suspension of aniline 2-6 (302 mg, 0.408 mmol, 1 equiv) andN-methyl-L-azetidine-2-carboxilic acid (148 mg, 1.28 mmol, 3.1 equiv) inCH₂Cl₂ (6 mL) was addedO-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate(395 mg, 1.23 mmol, 3 equiv) and DIEA (285 μL, 1.64 mmol, 4 equiv).After 16.5 h, the resulting orange solution was concentrated underreduced pressure and purified via preparative reverse phase HPLC on aWaters Autopurification system using a Sunfire Prep C18 OBD column [5μm, 19×50 mm; flow rate, 20 mL/min; Solvent A: H₂O with 0.1% HCO₂H;Solvent B: CH₃CN with 0.1% HCO₂H; injection volume: 4×2.5-3.0 mL(CH₃CN); gradient: 50→90% B over 15 min; mass-directed fractioncollection]. Two sets of fractions with the desired MW, eluting at4.6-6.5 min and 6.5-9.4 min, were collected separately and freeze-driedto provide 147 mg of 4-1-1 (43%): ¹H NMR (400 MHz, CDCl₃) δ 16.04 (s,1H), 10.10 (s, 1H), 8.48 (d, J=11.0 Hz, 1H), 7.54-7.48 (m, 4H),7.40-7.32 (m, 5H), 5.36 (s, 2H), 4.99 (d, J=9.8 Hz, 1H), 4.90 (d, J=9.8Hz, 1H), 3.96 (d, J=10.4 Hz, 1H), 3.54 (t, J=7.9 Hz, 1H), 3.39-3.34 (m,1H), 3.25-3.19 (m, 1H), 3.05-2.92 (m, 2H), 2.58-2.36 (m, 10H), 2.23-2.06(m, 4H), 0.81 (s, 9H), 0.28 (s, 3H), 0.11 (s, 3H); MS (ESI) m/z 837.37(M+H).

To a solution of 4-1-1 (147 mg, 0.175 mmol, 1 equiv) in dioxane (3.5 mL)was added an aqueous solution of HF (50%, 750 μL). After 4 h, thereaction solution was poured into an aqueous K₂HPO₄ solution (9 g in 90mL) and extracted with EtOAc (2×50 mL). The combined organic layers weredried (Na₂SO₄), filtered, and concentrated under reduced pressure toprovide 128.4 mg of a crude yellow foam.

The HF deprotection product (144 mg, 0.199 mmol, 1 equiv) was dissolvedin dioxane:MeOH (1:1, 4 mL), and palladium on carbon (10%, 43.5 mg) wasadded. The flask was fitted with a septum and evacuated and back-filledthree times with hydrogen gas. Hydrogen gas was bubbled through thereaction solution for three minutes, and the reaction mixture wasstirred under an atmosphere (balloon) of hydrogen gas for 3.25 h. Thereaction mixture was filtered through celite to remove the palladiumcatalyst and concentrated under reduced pressure. Preparative reversephase HPLC of this oil was performed on a Waters Autopurification systemusing a Polymerx 10μ RP-γ 100 R column [30×21.20 mm, 10 micron, solventA: 0.05 N HCl in water, solvent B: CH₃CN; injection volume: 2×3.2 mL(0.05 N HCl in water); gradient: 10→35% B over 20 min; mass-directedfraction collection]. Fractions with the desired MW, eluting at6.10-8.40 min and 6.9-9.4 min, respectively for each run, were combined.The pH of the solution at 0° C. was adjusted (pH 1.8 to pH 7.4) viadropwise addition of 0.5 M aqueous NaOH solution (approximately 7.8 mL)and careful monitoring with an electronic pH meter. The aqueous solutionwas extracted with CH₂Cl₂ (3×60 mL) and the combined organic layers weredried (Na₂SO₄), filtered, and concentrated under reduced pressure toprovide 79.7 mg of compound 89 as the free base (0.146 mmol, 73%). Thisyellow solid was dissolved in MeOH (3 mL), and MeSO₃H (19 μL, 0.292mmol, 2 equiv) was added. The solution was concentrated under reducedpressure, dried under vacuum, and freeze-dried from water to provide 105mg of 89 as the dimesylate salt. ¹H NMR (400 MHz, CD₃OD) δ 8.22 (d,J=11.0 Hz, 1H), 5.16 (t, J=8.6 Hz, 1H), 4.21-4.12 (m, 1H), 4.09-4.02 (m,2H), 3.17-2.85 (m, 10H), 2.68 (s, 6H, mesylate H), 2.64-2.59 (m, 1H),2.34-2.15 (m, 2H), 1.70-1.58 (m, 1H); MS (ESI) m/z 545.18 (M+H).

General Procedures for the Preparation of Compounds 90-94

To a solution of aniline 2-6 (1 equiv) in THF (0.05-0.09M) was added anacid chloride (3 equiv). The reaction solution was filtered throughcelite and concentrated under reduced pressure. The resulting oil wasdissolved in dioxane (1 mL) and an aqueous solution of HF (50%, 200 μL)was added. Upon completion, the reaction was poured into an aqueousK₂HPO₄ solution (2.6 g in 30 mL) and extracted with EtOAc (2×25 mL). Thecombined organic layers were dried (Na₂SO₄), filtered, and concentratedunder reduced pressure. Palladium on carbon (10%) was added to asolution of this crude oil in dioxane:MeOH (1:1, 1 mL). The flask wasfitted with a septum and evacuated and back-filled three times withhydrogen gas, and then the solution was degassed with bubbling hydrogenfor 2 minutes. The reaction was stirred under an atmosphere (balloon) ofhydrogen gas for 2 h. The reaction mixture was filtered through celiteto remove the palladium catalyst and concentrated under reducedpressure. The crude products were purified by preparative reverse phaseHPLC.

Prepared by above general procedure with the following reagents: aniline2-6 (21.1 mg, 0.028 mmol, 1 equiv), picolinoyl chloride hydrochloride(15.8 mg, 0.088, 3 equiv), with triethylamine (11.7 μL, 0.084 mmol, 3equiv), and 10% Pd-C (10 mg), provided a crude oil. Preparative reversephase HPLC of the crude product was performed on a WatersAutopurification system using a Polymerx 10μ RP-γ 100 R column [30×21.20mm, 10 micron, solvent A: 0.05 N HCl in water, solvent B: CH₃CN;injection volume: 2.5 mL (0.05 N HCl in water); gradient: 10→60% B over20 min; mass-directed fraction collection]. Fractions with the desiredMW, eluting at 14.8-16.4 min, were collected and freeze-dried to provide5.8 mg of the desired compound 90 (37%): ¹H NMR (400 MHz, CD₃OD) δ8.73-8.69 (m, 1H), 8.58-8.52 (m, 1H), 8.27-8.21 (m, 1H), 8.08-8.00 (m,1H), 7.66-7.60 (m, 1H), 4.09 (s, 1H), 3.29-2.92 (m, 9H), 2.38-2.18 (m,2H), 1.72-1.60 (m, 1H); MS (ESI) m/z 553.27 (M+H).

Prepared by above general procedure with the following reagents: aniline2-6 (31.0 mg, 0.042 mmol, 1 equiv), 1-methylpyrrole-2-carbonyl chloride(22 mg, 0.15 mmol, 3 equiv), and 10% Pd-C (10 mg). Preparative reversephase HPLC of the crude product was performed on a WatersAutopurification system using a Polymerx 10μ RP-γ 100 R column [30×21.20mm, 10 micron, solvent A: 0.05 N HCl in water, solvent B: CH₃CN;injection volume: 2.0 mL (0.05 N HCl in water); gradient: 20→70% B over20 min; mass-directed fraction collection]. Fractions with the desiredMW were collected and freeze-dried and repurified via the same systemwith the gradient: 10→60% B over 20 min. Fractions with the desired MW,eluting at 15.5-16.5 min, were collected and freeze-dried to provide 2.5mg of the desired compound 91 (11%): ¹H NMR (400 MHz, CD₃OD) δ 8.20 (d,J=11.6 Hz, 1H), 6.98-6.86 (m, 2H), 6.17-6.10 (m, 1H), 4.08 (s, 1H), 3.94(s, 3H), 3.19-2.90 (m, 9H), 2.33-2.18 (m, 2H), 1.80-1.56 (m, 1H); MS(ESI) m/z 555.32 (M+H).

Prepared by above general procedure with the following reagents: aniline2-6 (31.0 mg, 0.042 mmol, 1 equiv), 5-methylisoxazole-3-carbonylchloride (19.0 mg, 0.13 mmol, 3 equiv), and 10% Pd-C (10 mg).Preparative reverse phase HPLC of the crude product was performed on aWaters Autopurification system using a Polymerx 10μ RP-γ 100 R column[30×21.20 mm, 10 micron, solvent A: 0.05 N HCl in water, solvent B:CH₃CN; injection volume: 2.8 mL (0.05 N HCl in water); gradient: 10→60%B over 20 min; mass-directed fraction collection]. Fractions with thedesired MW, eluting at 14.5-15.5 min, were collected and freeze-dried toprovide 4.0 mg of the desired compound 92 (17%): ¹H NMR (400 MHz, CD₃OD)δ 8.32 (d, J=11.0 Hz, 1H), 6.59 (s, 1H), 4.09 (s, 1H), 3.19-2.90 (m,9H), 2.52 (s, 3H), 2.34-2.18 (m, 2H), 1.71-1.58 (m, 1H); MS (ESI) m/z557.26 (M+H).

Prepared by above general procedure with the following reagents: aniline2-6 (30.0 mg, 0.041 mmol, 1 equiv), 1-methyl-1H-pyrazole-3-carbonylchloride (16.8 mg, 0.12 mmol, 3 equiv), and 10% Pd-C (20 mg).Preparative reverse phase HPLC of the crude product was performed on aWaters Autopurification system using a Polymerx 10μ RP-γ 100 R column[30×21.20 mm, 10 micron, solvent A: 0.05 N HCl in water, solvent B:CH₃CN; injection volume: 3.2 mL (0.05 N HCl in water); gradient: 10→60%B over 20 min; mass-directed fraction collection]. Fractions with thedesired MW, eluting at 12.5-14.5 min, were collected and freeze-dried toprovide 11.2 mg of the desired compound 93 (49%): ¹H NMR (400 MHz,CD₃OD) δ 8.38 (d, J=11.0 Hz, 1H), 7.68 (s, 1H), 6.82-6.76 (m, 1H), 4.09(s, 1H), 3.99 (s, 3H), 3.16-2.90 (m, 9H), 2.31-2.16 (m, 2H), 1.70-1.56(m, 1H); MS (ESI) m/z 556.31 (M+H).

Prepared by above general procedure with the following reagents: aniline2-6 (30.0 mg, 0.041 mmol, 1 equiv), 1,3-thiazole-2-carbonyl chloride(17.8 mg, 0.12 mmol, 3 equiv), and 10% Pd-C (15 mg). Preparative reversephase HPLC of the crude product was performed on a WatersAutopurification system using a Polymerx 10μ RP-γ 100 R column [30×21.20mm, 10 micron, solvent A: 0.05 N HCl in water, solvent B: CH₃CN;injection volume: 3.2 mL (0.05 N HCl in water); gradient: 10→60% B over20 min; mass-directed fraction collection]. Fractions with the desiredMW, eluting at 14.6-17.0 min, were collected and freeze-dried to provide5.4 mg of the desired compound 94 (23%): ¹H NMR (400 MHz, CD₃OD) δ 8.38(d, J=11.0 Hz, 1H), 8.02 (d, J=3.0 Hz, 1H), 7.95 (d, J=2.4 Hz, 1H), 4.09(s, 1H), 3.20-2.90 (m, 9H), 2.34-2.17 (m, 2H), 1.70-1.56 (m, 1H); MS(ESI) m/z 559.23 (M+H).

Example 5. Synthesis of Compounds of Structural Formula (A), wherein Yis —N(R^(A))(R^(B)), or —NH—SO₂—(CH₂)₂—N(R^(A))(R^(B))

In Scheme 5, R represents —(C₁-C₆)alkyl, —(C₀-C₅)alkylene-carbocyclyl,—(C₀-C₅)alkylene-aryl, —(C₀-C₅)alkylene-heterocyclyl,—(C₀-C₅)alkylene-heteroaryl, —(C₁-C₃)alkylene-N(R^(A))(R^(B)); Arrepresents an aryl or a heteroaryl group; and R^(U) and R^(V) are R^(A)and R^(B), receptively, as defined in Structural Formula (B).

The following compounds were prepared according to Scheme 5.

Compound 2-6 (150 mg, 0.203 mmol, 1.0 equiv) was dissolved in1,2-dichloroethane (3 mL). HOAc (58.1 μL, 1.01 mmol, 5 equiv.) andisovaleraldehyde (32.9 μL, 0.304 mmol, 1.5 equiv) were added. Themixture was stirred for 1 h. Na(OAc)₃BH (129 mg, 0.609 mmol, 3.0 equiv)was added and the resulting mixture was stirred for another hour. Themixture was washed with H₂O (10 mL) and concentrated to give crude 5-1-1(250 mg), which was used for the next step without further purification:MS (ESI) m/z 810.59 (M+H).

Aqueous HF (0.3 mL, 48-50%) was added to a CH₃CN solution (1.5 mL) of5-1-1 (250 mg crude) in a plastic vial at 25° C. The reaction wasstirred at 25° C. for 18 hrs. The resulting mixture was poured into anaqueous solution (10 mL) of K₂HPO₄ (2 g). The solution was extractedwith EtOAc (3×15 mL). The combined EtOAc extracts were dried over sodiumsulfate and concentrated to give the crude intermediate (155 mg).

10% Pd-C (20 mg) was added to a dioxane/MeOH solution (4 mL, 1:1) of theabove crude intermediate. HCl/MeOH (0.5 mL, 0.5 N) was also added. Thereaction mixture was stirred under H₂ (balloon) at 25° C. for 2 hrs andfiltered through a pad of Celite. The filtrate was concentrated to givethe crude product 144 mg. The crude product was purified by HPLC on aPolymerx 10μ RP-γ 100 R column [30×21.20 mm, 10 micron, solvent A: 0.05N HCl, solvent B: CH₃CN, sample in 2.0 mL (0.05 N HCl), gradient elutionwith 10→100% B over 15 min, mass-directed fraction collection] to yieldthe desired product 95 as a yellow solid (82 mg, 78%, 2 steps): ¹H NMR(400 MHz, CD₃OD) δ 7.44 (d, J=9.2 Hz, 1H), 4.12 (s, 1H), 3.42-3.37 (m,2H), 3.05 (s, 3H), 2.97 (s, 3H), 3.21-2.97 (m, 3H), 2.39-2.30 (m, 1H),2.29-2.22 (m, 1H), 1.79-1.59 (m, 4H), 0.98 (d, J=6.4 Hz, 6H); MS (ESI)m/z 518.43 (M+H).

Compounds 96-101 were prepared similarly to compound 95 using thecorresponding aldehydes in the reductive alkylation step.

¹H NMR (400 MHz, CD₃OD) δ 7.39 (d, J=9.2 Hz, 1H), 4.10 (s, 1H), 3.34 (t,J=7.8 Hz, 2H), 3.04 (s, 3H), 2.96 (s, 3H), 3.21-2.95 (m, 3H), 2.35 (t,J=13.7 Hz, 1H), 2.27-2.20 (m, 1H), 1.82-1.72 (m, 2H), 1.71-1.60 (m, 1H),1.05 (t, J=7.4 Hz, 3H); MS (ESI) m/z 490.32 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 7.34 (d, J=9.2 Hz, 1H), 4.10 (s, 1H), 3.34 (t,J=7.8 Hz, 2H), 3.04 (s, 3H), 2.96 (s, 3H), 3.24-2.95 (m, 11H), 2.33 (t,J=13.7 Hz, 1H), 2.27-2.20 (m, 1H), 2.11-1.98 (m, 1H), 1.71-1.60 (m, 1H),1.08 (d, J=6.9 Hz, 6H); MS (ESI) m/z 504.46 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 7.43 (d, J=8.7 Hz, 1H), 4.10 (s, 1H), 3.34 (t,J=7.8 Hz, 2H), 3.04 (s, 3H), 2.96 (s, 3H), 3.28-2.95 (m, 11H), 2.41-2.31(m, 1H), 2.27-2.20 (m, 1H), 2.11-1.98 (m, 1H), 1.72-1.60 (m, 1H),1.20-1.11 (m, 1H), 0.74-0.68 (m, 2H), 0.43-0.38 (m, 2H); MS (ESI) m/z502.40 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 6.97-6.89 (m, 1H), 4.07 (s, 1H), 3.34 (t,J=7.8 Hz, 2H), 3.03 (s, 3H), 2.95 (s, 3H), 3.14-2.92 (m, 11H), 2.30-2.15(m, 2H), 1.89-1.59 (m, 7H), 1.38-1.20 (m, 3H), 1.11-1.00 (m, 2H); MS(ESI) m/z 544.50 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 6.83 (d, J=10.5 Hz, 1H), 4.06 (s, 1H), 3.34(t, J=7.8 Hz, 2H), 3.03 (s, 3H), 2.95 (s, 3H), 3.11-2.93 (m, 5H),2.27-2.14 (m, 2H), 1.67-1.57 (m, 1H), 1.04 (s, 9H); MS (ESI) m/z 518.48(M+H).

¹H NMR (400 MHz, CD₃OD) δ 7.46-7.42 (m, 1H), 4.15 (s, 1H), 3.33 (s, 6H),3.04 (s, 3H), 2.96 (s, 3H), 3.17-2.95 (m, 3H), 2.44-2.34 (m, 1H),2.29-2.22 (m, 1H), 1.71-1.60 (m, 1H); MS (ESI) m/z 476.29 (M+H).

Prepared similarly to 95 using tBuN(Cbz)CH₂CHO: ¹H NMR (400 MHz, CD₃OD)δ 6.72 (d, J=11.0 Hz, 1H), 4.07 (s, 1H), 3.54-3.46 (m, 2H), 3.26-3.19(m, 2H), 3.03 (s, 3H), 2.95 (s, 3H), 3.14-2.92 (m, 3H), 2.23-2.14 (m,2H), 1.67-1.55 (m, 1H), 1.38 (s, 9H); MS (ESI) m/z 547.51 (M+H).

Compound 103 was also isolated from the preparation of 102: ¹H NMR (400MHz, CD₃OD) δ 6.71 (d, J=11.0 Hz, 1H), 4.07 (s, 1H), 3.47 (t, J=6.0 Hz,2H), 3.17 (t, J=6.0 Hz, 2H), 3.03 (s, 3H), 2.95 (s, 3H), 3.13-2.92 (m,3H), 2.23-2.12 (m, 2H), 1.66-1.54 (m, 1H); MS (ESI) m/z 491.42 (M+H).

A vessel containing aniline 2-6 (18.2 mg, 0.024 mmol, 1 equiv), Pd₂dba₃(3.0 mg, 0.0033 mmol, 0.13 equiv), Xantphos (3.4 mg, 0.0059 mmol, 0.25equiv), K₃PO₄ (40 mg, 0.188 mmol, 7.8 equiv) and 4,6-dichloropyrimidine(6.5 mg, 0.044 mmol, 1.8 equiv) was evacuated and back-filled withnitrogen gas three times. Dioxane (500 μL) was added, and the reactionmixture stirred vigorously and heated at 80° C. for 4.5 h. The reactionmixture was filtered through celite and concentrated under reducedpressure. Preparative reverse phase HPLC of the resulting yellow oil wasperformed on a Waters Autopurification system using a Sunfire Prep C18OBD column [5 μm, 19×50 mm; flow rate, 20 mL/min; Solvent A: H₂O with0.1% HCO₂H; Solvent B: CH₃CN with 0.1% HCO₂H; injection volume: 1.8 mL(CH₃CN); gradient: 80→100% B over 15 min; mass-directed fractioncollection]. Fractions with the desired MW, eluting at 9.2-9.8 min, werecollected and freeze-dried to provide 7.5 mg of compound 5-3-1 (37%). ¹HNMR (400 MHz, CDCl₃) δ 15.97 (s, 1H), 8.48 (s, 1H), 8.33 (d, J=5.5 Hz,1H), 7.52-7.46 (m, 2H), 7.40-7.28 (m, 8H), 7.07 (s, 1H), 6.11 (s, 1H),5.34 (s, 2H), 4.97 (d, J=11.6 Hz, 1H0, 4.88 (d, J=11.0 Hz, 1H), 3.95 (d,J=10.4 Hz, 1H), 3.28-3.19 (m, 1H), 3.09-2.98 (m, 1H), 2.61-2.54 (m, 1H),2.54-2.39 (m, 8H), 2.16 (d, J=14.6 Hz, 1H), 0.83 (s, 9H), 0.28 (s, 3H),0.14 (s, 3H); MS (ESI) m/z 852.57 (M+H).

To a solution of 5-3-1 (7.5 mg, 0.0088 mmol, 1 equiv) in dioxane (1.4mL) was added an aqueous solution of HF (50%, 200 μL). After 15.5 h, thereaction solution was poured into an aqueous K₂HPO₄ solution (2.4 g in20 mL) and extracted with EtOAc (2×20 mL). The combined organic layerswere dried (Na₂SO₄), filtered, and concentrated under reduced pressure.Palladium on carbon (10%, 10 mg) was added to a solution of this oil indioxane:MeOH (1:1, 1 mL). The flask was fitted with a septum andevacuated and back-filled three times with hydrogen gas. Hydrogen gaswas bubbled through the reaction solution for three minutes, and thereaction mixture was stirred under an atmosphere (balloon) of hydrogengas for 2.5 h. The reaction mixture was filtered through celite toremove the palladium catalyst and concentrated under reduced pressure.Preparative reverse phase HPLC purification of the resulting oil wasperformed on a Waters Autopurification system using a Polymerx 10μ RP-γ100 R column [30×21.20 mm, 10 micron, solvent A: 0.05 N HCl in water,solvent B: CH₃CN; injection volume: 2.0 mL (0.05 N HCl in water);gradient elution with 10→50% B over 10 min, then held at 100% for 5 min;mass-directed fraction collection]. Fractions with the desired MW,eluting at 6.90-7.80 min, were collected and freeze-dried to provide 2.2mg of 104 (48%). ¹H NMR (400 MHz, CD₃OD) δ 8.83 (s, 1H), 8.37-8.25 (m,1H), 8.18-8.05 (m, 1H), 7.30-7.20 (m, 1H), 4.10 (s, 1H), 3.20-2.90 (m,9H), 2.40-2.29 (m, 1H), 2.27-2.19 (m, 1H), 1.72-1.58 (m, 1H); MS (ESI)m/z 526.31 (M+H).

To a solution of aniline 2-6 (30.0 mg, 0.041 mmol, 1 equiv) in1,2-dichloroethane (500 μL) was added pyridine (16.3 μL, 0.20 mmol, 5equiv) and 2-chloroethanesulfonyl chloride (21 μL, 0.20 mmol, 5 equiv).The reaction vessel was sealed and heated to 45° C. After one hour, thereaction was a solid yellow gel, and another 500 μL 1,2-dichloroethanewas added to form a suspension and the reaction was sealed and heated to45° C. After 18.5 h pyrrolidine (68 μL, 0.82 mmol, 20 equiv) was addedand the reaction heated to 45° C. for 2.5 hours. The solution was pouredinto aqueous pH 7 phosphate buffer (8 mL) and extracted with EtOAc (2×25mL). The combined organic layers were dried (Na₂SO₄), filtered, andconcentrated under reduced pressure. To a solution of this crude oil inCH₃CN (1.8 mL) was added an aqueous solution of HF (50%, 300 μL). After15 h, the reaction solution was poured into an aqueous K₂HPO₄ solution(3.6 g in 30 mL) and extracted with EtOAc (3×15 mL). The combinedorganic layers were dried (Na₂SO₄), filtered, and concentrated underreduced pressure. Palladium on carbon (10%, 8.4 mg) was added to asolution of this oil in dioxane:MeOH (1:1, 1.2 mL). The flask was fittedwith a septum and evacuated and back-filled three times with hydrogengas, and the reaction mixture was stirred under an atmosphere (balloon)of hydrogen gas for 1.5 h. Another 10 mg palladium catalyst was addedand the reaction was evacuated and back-filled with hydrogen gas asbefore. After 6 h, the reaction mixture was filtered through celite toremove the palladium catalyst and concentrated under reduced pressure.Preparative reverse phase HPLC purification of the resulting oil wasperformed on a Waters Autopurification system using a Polymerx 10 μRP-γ100 R column [30×21.20 mm, 10 micron, solvent A: 0.05 N HCl in water,solvent B: CH₃CN; injection volume: 3.5 mL (0.05 N HCl in water);gradient elution with 0→100% B over 10 min, then held at 100% for 5 min;mass-directed fraction collection]. Two sets of fractions with thedesired MW, eluting at 6.3-7.1 min and 8.7-9.3 min, were collectedseparately and freeze-dried to provide 9.7 mg of crude compounds 105.Purification via preparative reverse phase HPLC with gradient elutionwith 20→70% B over 20 min; mass-directed fraction collection] provided3.3 mg of pure 105 (13%): ¹H NMR (400 MHz, CD₃OD) δ 7.44 (d, J=9.8 Hz,1H), 4.09 (s, 1H), 3.79-3.65 (m, 4H), 3.63-3.56 (m, 2H), 3.18-2.90 (m,11H), 2.35-2.26 (m, 1H), 2.26-2.10 (m, 3H), 2.10-1.96 (m, 2H), 1.69-1.59(m, 1H); MS (ESI) m/z 609.36 (M+H).

Compound 106 (1.7 mg, 7%) was also isolated from the preparation ofcompound 105: ¹H NMR (400 MHz, CD₃OD) δ 6.71 (d, J=11.0 Hz, 1H), 4.06(s, 1H), 3.67-3.60 (m, 2H), 3.38-3.33 (m, 4H), 3.09-2.90 (m, 9H),2.24-2.13 (m, 2H), 1.95-1.91 (m, 5H), 1.90-1.85 (m, 1H), 1.68-1.55 (m,1H); MS (ESI) m/z 609.36 (M+H).

Example 6. Synthesis of Compounds 107 and 108

The following compounds were prepared according to Scheme 6.

Compound 2-3 (5.0 g, 14.25 mmol, 1.0 equiv) in MeOH (20 mL) was added aaqueous solution of HCHO (2.3 g, 37%, 28.50 mmol, 2.0 equiv) andpalladium on carbon (0.5 g, 10 wt %). The reaction was purged withhydrogen and stirred under H₂ (balloon) at room temperature for 2 hours.The reaction mixture was filtered through celite and concentrated toafford 1.3 g crude compound 6-1 as a yellow solid.

To compound 6-1 (0.9 g, 3.27 mmol, 1.0 equiv) in DCM was added Boc₂O(2.14 g, 9.81 mmol, 3.0 equiv) dropwise. DMAP (135 mg, 15 wt %) wasadded to the mixture and the reaction was stirred at room temperaturefor 1 hour. Then the reaction mixture was heated to reflux for 1 hour.The reaction mixture was concentrated. The crude compound was purifiedby column chromatography on silica gel eluted with (PE:EA=200:1→100:1)to yield compound 6-2 (1.16 g, 73.4%) as a light yellow solid.

¹H NMR (400 MHz, DMSO) δ 7.60 (d, J=10.0 Hz, 1H), 7.53-7.44 (m, 2H),7.36-7.31 (m, 1H), 7.28-7.22 (m, 2H), 3.06 (s, 3H), 2.33 (d, J=2.0 Hz,3H), 1.38 (s, 9H), 1.34 (s, 9H); MS (ESI) m/z 476.2 (M+H).

To diisopropylamine (0.28 mL, 3.2 mmol, 10.0 equiv) in THF at −78° C.was added nBuLi (0.8 mL, 2.50 M/hexane, 3.2 mmol, 10.0 equiv) and TMEDA(0.40 mL, 5.0 mmol, 10.0 equiv) at −78 C dropwise. The reaction wasstirred at −78 C for 40 min. Compound 6-2 (480 mg, 1.0 mmol, 3.0 equiv)in THF was added to the reaction mixture dropwise at −78 C. Theresulting deep-red solution was stirred at −78° C. 60 min, the enone(160 mg 0.33 mmol, 1.0 equiv) in THF was added to the mixture dropwiseat −78 C. The deep-red solution was gradually warmed up with stirringfrom −78 C to −20° C. over a period of 1 h. The resulting orangesolution was brought to 0° C., and quenched with aqueous saturatedammonium chloride (100 mL). The yellow-green mixture was extracted withEtOAc two times. The combined EtOAc extracts were dried (Na₂SO₄) andconcentrated to yield the crude product. Flash column chromatography onsilica gel with 0%, 5%, 10%, EtOAc/hexane sequentially yielded thedesired product 6-3 as a light-yellow solid (42 mg, 14.8%). ¹H NMR (400MHz, CDCl₃) δ 15.70 (s, 1H), 7.52-7.50 (m, 2H), 7.42-7.33 (m, 3H), 7.16(d, J=8.4 Hz, 1H), 5.37 (s, 2H), 3.95 (d, J=10.8 Hz, 1H), 3.28-3.23 (m,1H), 3.14 (s, 3H), 3.10-3.05 (m, 1H), 2.58-2.47 (m, 9H), 2.16 (d, J=14.0Hz, 1H), 1.53 (s, 9H), 1.42 (s, 9H), 0.89 (s, 9H), 0.29 (s, 3H), 0.15(s, 3H); MS (ESI) m/z 864.43 (M+H).

Compound 6-3 (120 mg, 0.14 mmol) was dissolved in THF (5 mL) and aqueousHF (40%, 2 mL) was added dropwise. The yellow solution was stirred atroom temperature overnight. The resulting deep-red solution was slowlyadded into an aqueous K₂HPO₄ solution with stirring. The pH of themixture was adjusted by aqueous K₂HPO₄ solution to about 8. The yellowmixture was extracted with EtOAc two times. The combined EtOAc extractswere dried (Na₂SO₄) and concentrated to yield the crude product.

The above crude compound (120 mg, crude, ˜0.14 mmol, 1.0 equiv) wasdissolved in HPLC grade MeOH (10 mL) and 10% Pd—C (25 mg, 0.03 mmol, 0.2equiv) were added. The mixture was purged with hydrogen by bubblinghydrogen through with gentle stirring for 5 min. The reaction was thenvigorously stirred under hydrogen balloon at room temperature for 2 hr.LC-MS analysis indicated the reaction completed. The catalyst wasfiltered off and the mixture was concentrated, the residue was purifiedby reverse phase HPLC to afford the desired compound 107 (50 mg, 78%) asa yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 7.462 (d, J=8.4 Hz, 1H), 4.14(s, 1H), 3.21-2.93 (m, 9H), 3.10 (s, 3H), 2.38-2.25 (m, 2H), 1.68-1.62(m, 1H); MS (ESI) m/z 462.2 (M+H).

Compound 107 (15 mg, 0.033 mmol, 1.0 equiv) in THF (2 mL) was added withpyrrolidin-1-yl-acetic acid (10.2 mg, 0.066 mmol, 2.0 equiv), Na₂CO₃(10.2 mg, 0.066 mmol, 2.0 equiv) and HATU (25.5 mg, 0.066 mmol, 2.0equiv). The reaction mixture was stirred at room temperature for 48hours. LC-MS analysis indicated the reaction completed. The reactionmixture was concentrated under vacuum, the crude product was purified byreverse phase HPLC to afford the desired compound 108 (2.1 mg) as ayellow solid. ¹H NMR (400 MHz, CD₃OD) δ 7.44-7.40 (m, 1H), 4.02-3.97 (m,2H), 3.83-3.76 (m, 1H), 3.60-3.58 (m, 2H), 3.15 (d, J=6.4 Hz, 3H),3.03-2.83 (m, 11H), 2.31-2.13 (m, 2H), 2.03-1.85 (m, 4H), 1.61-1.52 (m,1H); MS (ESI) m/z 572.9 (M+H).

Example 7. Synthesis of Compounds 109-112

The following compounds were prepared according to Scheme 7.

To Boc-L-glutamic acid-1-benzyl ester (7-1) (3.00 g, 8.89 mmol, 1.0 eq)in DMF (20 mL) at rt was added potassium carbonate (1.84 g, 13.33 mmol,1.5 eq) and methyl iodide (0.67 mL, 10.74 mmol, 1.2 eq). The mixture wasdiluted with EtOAc (200 mL), washed with water (200 mL), saturatedaqueous sodium bicarbonate (100 mL×2), and brine (100 mL×1). The EtOAcsolution was dried over sodium sulfate and concentrated in vacuo: R_(f)0.33 (20% EtOAc/hexane).

Boc₂O (2.91 g, 13.33 mmol, 1.5 eq), DMAP (54 mg, 0.44 mmol, 0.05 eq),and DIEA (3.10 mL, 17.80 mmol, 2 eq) were added to the aboveintermediate in acetonitrile (20 mL). The solution was stirred at rt for60 hrs, added with saturated aqueous sodium bicarbonate (100 mL), andextracted with EtOAc (100 mL x 1, 50 mL x 2). The EtOAc extracts werecombined, dried over sodium sulfate, and concentrated in vacuo to yieldthe desired product 7-2 as a pale liquid (quantitative): R_(f) 0.45 (20%EtOAc/hexane); ¹H NMR (400 MHz, CDCl₃) δ 7.25-7.35 (m, 5H), 5.14 (s,2H), 4.95 (dd, J=4.9, 9.8 Hz, 1H), 3.65 (s, 3H), 2.43-2.52 (m, 1H),2.37-2.42 (m, 2H), 2.15-2.25 (m, 1H), 1.42 (s, 18H); MS (ESI) m/z 452.3(M+H).

To compound 7-2 (8.89 mmol, 1 eq) in anhydrous diethyl ether (40 mL) at−78° C. was added DIBAL-H (12.33 mL, 1 M/hexane, 12.33 mmol, 1.25 eq)dropwise. The reaction was stirred at −78° C. for 2 hrs. AdditionalDIBAL-H (1.20 mL, 1 M/hexane, 1.20 mmol) was added. The reaction wasstirred at −78° C. for another 1 hr and quenched with HOAc (2.80 mL) at−78° C. The reaction was warmed to rt and added with 10% aqueous sodiumcarbonate (75 mL). The mixture was stirred for 15 min and extracted withmethylene chloride (200 mL x 1, 50 mL x 2). The methylene chlorideextracts were combined, dried over sodium sulfate, and concentrated invacuo to yield the desired product 7-3 (quantitative): R_(f) 0.40 (20%EtOAc/hexane); ¹H NMR (400 MHz, CDCl₃) δ 9.75 (s, 1H), 7.25-7.35 (m,5H), 5.14 (s, 2H), 4.87-4.92 (m, 1H), 2.45-2.65 (m, 3H), 2.12-2.22 (m,1H), 1.42 (s, 18H); MS (ESI) m/z 422.3 (M+H).

To aniline 9 (90 mg, 0.20 mmol, bis-HCl salt, 1 eq) in anhydrous DMF (2mL) was added aldehyde 7-3 (101 mg, 0.24 mmol, 1.2 eq), triethylamine(0.028 mL, 0.20 mmol, 1 eq), and Na(OAc)₃BH (64 mg, 0.30 mmol, 1.5 eq).The solution was stirred at rt for 1 hr and added slowly into diethylether (50 mL) with rapid stirring. The yellow solid was collected,washed with more diethyl ether (5 mL×3), and dried under vacuum toafford the intermediate 7-4.

Intermediate 7-4 was dissolved in dioxane/methanol (5 mL, 1:4 v/v,containing 0.1 N HCl). 10% Pd—C (85 mg, 0.04 mmol, 0.2 eq) was added.The mixture was purged with hydrogen and stirred under 1 atm hydrogen atrt for 1 hr. The catalyst was filtered with a small Celite pad andwashed with methanol (2 mL×3). The filtrate was concentrated in vacuo.The crude product was purified by reverse phase preparative HPLC usingmethanol and 0.05 N HCl/water as mobile phases. Free-drying yieldedmostly the Boc-deprotected product as a brown solid (25 mg, 22%, 2steps), which was re-protected by treatment with Boc₂O (11 mg, 0.050mmol, 1.1 eq) and DIEA (0.039 mL, 0.22 mmol, 5 eq) in THF/water (5 mL,1:1 v/v) at rt for 1 hr. Concentration yielded the desired product 7-5as a yellow solid: MS (ESI) m/z 663.2 (M+H), which was used directly inthe subsequent steps without further purification.

To a suspension of compound 7-5 (0.044 mmol, 1 eq) and sodium carbonate(7 mg, 0.066 mmol, 1.5 eq) in THF at rt was added HATU (20 mg, 0.053mmol, 1.2 eq). The mixture was rapidly stirred at rt for 2 hrs. Methanol(5 mL) was added. The solids were filtered off. The filtrate wasconcentrated under reduced pressure to yield crude 7-6 as a yellowsolid: MS (ESI) m/z 645.1 (M+H).

Compound 7-6 (0.044 mmol) was treated with 4 N HCl/dioxane (5 mL) at rtfor overnight and concentrated in vacuo. The residue was re-dissolved inmethanol (1 mL) and added dropwise into diethyl ether (50 mL) with rapidstirring. The yellow precipitates were collected, washed with morediethyl ether (5 mL×3), and dried under vacuo to afford crude 109 as abrown solid.

One fifth of the above crude product was purified by reverse phase HPLCto yield pure 109 as a yellow solid (1.5 mg, 31%): ¹H NMR (400 MHz,CD₃OD) δ 7.36 (d, 9.2 Hz, 1H), 4.09-4.15 (m, 1H), 4.08 (s, 1H),3.70-3.80 (m, 1H), 3.58-3.68 (m, 1H), 2.90-3.50 (m, 12H), 2.30-2.45 (m,2H), 2.10-2.25 (m, 3H), 1.95-2.10 (m, 1H), 1.58-1.70 (m, 1H); MS (ESI)m/z 545.1 (M+H).

To 2/5 of crude 109 (0.018 mmol, 1 eq) in DMF (1 mL) was added aqueousformaldehyde (0.007 mL, 36.5%/water, 0.094 mmol, 5 eq), InCl₃ (0.4 mg,0.002 mmol, 0.1 eq), and Na(OAc)₃BH (15 mg, 0.071 mmol, 4 eq). Thereaction was stirred at rt for 2 hrs and quenched with 0.5 NHCl/methanol (1 mL). The solution was added dropwise into diethyl ether(100 mL) with rapid stirring. The precipitates were collected, washedwith more diethyl ether (2 mL×4), and purified by reverse phase HPLC toafford the desired compound 110 as a yellow solid (1.8 mg, 18%): ¹H NMR(400 MHz, CD₃OD) δ 7.7.44 (d, J=9.1 Hz, 1H), 4.37 (dd, J=6.1, 11.6 Hz,1H), 4.09 (s, 1H), 3.60-3.75 (m, 2H), 2.92-3.50 (m, 15H), 2.86 (s, 3H),2.10-2.50 (m, 6H), 1.60-1.72 (m, 1H); MS (ESI) m/z 573.3 (M+H).

To 2/5 of crude 109 (0.018 mmol, 1 eq) in DMF (1 mL) was addedcyclopropanecarboxaldehyde (1.4 μL, 0.018 mmol, 1 eq), InCl₃ (0.4 mg,0.002 mmol, 0.1 eq), and Na(OAc)₃BH (6 mg, 0.028 mmol, 1.5 eq). Thereaction was stirred at rt for overnight and quenched with 0.5 NHCl/methanol (1 mL). The solution was added dropwise into diethyl ether(100 mL) with rapid stirring. The precipitates were collected, washedwith more diethyl ether (2 mL×4), and purified by reverse phase HPLC toafford the desired compound 111 as a yellow solid (1.3 mg, 12%): ¹H NMR(400 MHz, CD₃OD) δ 7.38 (d, J=9.2 Hz, 1H), 4.22 (dd, J=6.1, 11.6 Hz,1H), 4.09 (d, 1H), 3.60-3.78 (m, 2H), 2.85-3.50 (m, 12H), 2.00-2.50 (m,6H), 1.60-1.72 (m, 1H), 1.10-1.20 (m, 1H), 0.70-0.75 (m, 2H), 0.40-0.50(m, 2H); MS (ESI) m/z 599.4 (M+H).

Dialkylated product 112 was also isolated from the preparation ofcompound 111 (1.0 mg, yellow solid, 9%): ¹H NMR (400 MHz, CD₃OD) δ 7.42(d, J=9.2 Hz, 1H), 4.70-4.80 (m, 1H), 4.09 (s, 1H), 3.55-3.80 (m, 3H),2.95-3.50 (m, 13H), 2.10-2.50 (m, 6H), 1.55-1.75 (m, 1H), 1.20-1.30 (m,2H), 0.68-0.90 (m, 4H), 0.38-0.58 (m, 4H); MS (ESI) m/z 653.3 (M+H).

Example 8. Synthesis of Compounds of Structural Formula (A), wherein Yis —(C₁-C₄)alkylene-N(R^(A))(R^(B)), or—(C₁-C₄)alkylene-N(R^(F))—C(O)—[C(R^(D))(R^(E))]₀₋₄—N(R^(A))(R^(B))

In Scheme 8, R and R′ is R^(B) and R^(A), respectively, as defined inStructural Formula (A) and R^(w) represents—[C(R^(D))(R^(E))]₁₋₄—N(R^(A))(R^(B)).

The following compounds were prepared according to Scheme 8.

Benzyl N-(hydroxymethyl)carbamate (92 mg, 0.51 mmol, 2.0 equiv) wasadded to a TFA/CH₃SO₃H (1 mL/1 mL) solution of compound 7 (110 mg, 0.25mmol) at 25° C. The reaction was stirred at 25° C. for 30 min.Preparative reverse phase HPLC purification on a Waters Autopurificationsystem using a Phenomenex Polymerx 10μ RP-1 100 A column [10 μm,150×21.20 mm; flow rate, 20 mL/min; Solvent A: 0.05 N HCl; Solvent B:CH₃CN; injection volume: 4.0 mL (0.05 N HCl/water); gradient: 0→30% Bover 20 min; mass-directed fraction collection]. Fractions with thedesired MW were collected and freeze-dried to yield 23 mg of pure 113:¹H NMR (400 MHz, CD₃OD) δ 7.47 (d, J=9.2 Hz, 1H), 4.16 (s, 2H), 4.13 (s,1H), 3.21-2.94 (m, 3H), 3.06 (s, 3H), 2.97 (s, 3), 2.37-2.22 (m, 2H),1.70-1.58 (m, 1H); MS (ESI) m/z 462.26 (M+H).

Et₃N (2 μL, 0.0136 mmol, 2.0 equiv) was added to a mixture of 113 (3 mg,0.0065 mmol) and pivaldehyde (0.8 μL, 0.00715 mmol, 1.1 equiv) in DMF(0.1 mL) at 25° C. The reaction was stirred at 25° C. for 15 min.NaBH(OAc)₃ (3 mg, 0.013 mmol) and HOAc (2 μL) was added to the resultingmixture. The reaction was stirred at 25° C. for 1 h. Preparative reversephase HPLC purification on a Waters Autopurification system using aPhenomenex Polymerx 10μ RP-1 100 A column [10 μm, 150×21.20 mm; flowrate, 20 mL/min; Solvent A: 0.05 N HCl; Solvent B: CH₃CN; injectionvolume: 4.0 mL (0.05 N HCl/water); gradient: 0→100% B over 15 min;mass-directed fraction collection]. Fractions with the desired MW werecollected and freeze-dried to yield 1 mg of 114: ¹H NMR (400 MHz, CD₃OD)δ 7.52 (d, J=9.1 Hz, 1H), 4.30 (s, 2H), 4.09 (s, 1H), 3.23-2.93 (m, 5H),3.04 (s, 3H), 2.95 (s, 3H), 2.40-2.19 (m, 2H), 1.71-1.60 (m, 1H), 1.05(s, 9H); MS (ESI) m/z 532.27 (M+H).

Compounds 115-118 were prepared similarly to compound 114 using thecorresponding aldehydes.

¹H NMR (400 MHz, CD₃OD) δ 7.51 (d, J=8.8 Hz, 1H), 4.25 (s, 2H), 4.10 (s,1H), 3.25-2.90 (m, 5H), 3.05 (s, 3H), 2.96 (s, 3H), 2.40-2.21 (m, 2H),1.90-1.60 (m, 7H), 1.42-0.95 (m, 5H); MS (ESI) m/z 558.31 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 7.50 (d, J=9.0 Hz, 1H), 4.24 (s, 2H), 4.09 (s,1H), 3.25-2.90 (m, 5H), 3.07 (s, 3H), 2.94 (s, 3H), 2.40-2.21 (m, 2H),1.82-1.58 (m, 3H), 1.01 (t, J=6.7 Hz, 3H); MS (ESI) m/z 504.22 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 7.51 (d, J=8.9 Hz, 1H), 4.23 (s, 2H), 4.09 (s,1H), 3.25-2.92 (m, 4), 3.02 (s, 3H), 2.95 (s, 3H), 2.40-2.19 (m, 2H),1.71-1.60 (m, 1H), 1.40 (d, J=7.0 Hz, 6H); MS (ESI) m/z 504.23 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 7.54 (d, J=9.1 Hz, 1H), 4.37 (s, 2H), 4.10 (s,1H), 3.20-2.85 (m, 3H), 3.05 (s, 3H), 2.97 (s, 3H), 2.91 (s, 3H), 2.90(s, 3H), 2.42-2.20 (m, 2H), 1.72-1.60 (m, 1H); MS (ESI) m/z 490.19(M+H).

Prepared from compound 114 by reductive alkylation using formaldehydeunder similar conditions: ¹H NMR (400 MHz, CD₃OD) δ 7.57 (d, J=9.1 Hz,1H), 4.61 (d, J=12.8 Hz, 1H), 4.27 (dd, J=12.8, 6.4 Hz, 1H), 4.10 (s,1H), 3.25-2.90 (m, 5), 3.03 (s, 3H), 2.96 (s, 3H), 2.95 (s, 3H),2.42-2.33 (m, 1H), 2.29-2.20 (m, 1H), 1.72-1.61 (m, 1H), 1.10 (d, J=6Hz, 9H); MS (ESI) m/z 546.30 (M+H).

Prepared similarly to 114 by reductive alkylation of 113 witht-Bu-N(Cbz)-CH₂CHO followed by hydrogenation: ¹H NMR (400 MHz, CD₃OD) δ7.59 (d, J=8.6 Hz, 1H), 4.38 (s, 2H), 4.09 (s, 1H), 3.60-2.95 (m, 7H),3.03 (s, 3H), 2.96 (s, 3H), 2.41-2.30 (m, 1H), 2.28-2.20 (m, 1H),1.72-1.60 (m, 1H), 1.44 (s, 9H); MS (ESI) m/z 561.31 (M+H).

2-t-Butylaminoacetylchloride hydrochloride (5.8 mg, 0.031 mmol, 1.2equiv) was added to a DMF solution (0.2 mL) of 113 (12 mg, 0.026 mmol)at 25° C. The reaction was stirred at 25° C. for 30 min. The reactionmixture was diluted with 0.05 N HCl (2 mL) and injected into a WatersAutopurification system equipped with a Phenomenex Polymerx 10μ RP-1 100A column [10 μm, 150×21.20 mm; flow rate, 20 mL/min; Solvent A: 0.05 NHCl; Solvent B: CH₃CN; gradient: 0→100% B over 15 min; mass-directedfraction collection]. Fractions with the desired MW were collected andfreeze-dried to yield 3.0 mg of pure 121: ¹H NMR (400 MHz, CD₃OD) δ 7.34(d, J=9.6 Hz, 1H), 4.46 (s, 2H), 4.08 (s, 1H), 3.81 (s, 2H), 3.18-2.92(m, 3H), 3.03 (s, 3H), 2.96 (s, 3H), 2.32-2.18 (m, 2H), 1.69-1.60 (m,1H), 1.38 (s, 9H); MS (ESI) m/z 575.30 (M+H).

Prepared similarly to compound 121: ¹H NMR (400 MHz, CD₃OD) δ 7.33 (d,J=9.9 Hz, 1H), 4.46 (s, 2H), 4.08 (s, 1H), 4.00 (s, 2H), 3.23-2.91 (m,3), 3.04 (s, 3H), 2.97 (s, 3H), 2.95 (s, 6H), 2.32-2.18 (m, 2H),1.70-1.58 (m, 1H); MS (ESI) m/z 547.23 (M+H).

Prepared similarly to 121 using n-propyl isocyanate: ¹H NMR (400 MHz,CD₃OD) δ 7.24 (d, J=9.8 Hz, 1H), 4.31 (s, 2H), 4.08 (s, 1H), 3.18-2.93(m, 3H), 3.10 (t, J=6.7 Hz, 2H), 3.03 (s, 3H), 2.96 (s, 3H), 2.32-2.18(m, 2H), 1.69-1.58 (m, 1H), 1.55-1.46 (m, 2H), 0.92 (t, J=6.7 Hz, 3H);MS (ESI) m/z 584.01 (M+H).

Example 9. Synthesis of Compounds of Structural Formula (A), wherein Yis —(CH₂)₃—N(R^(A))(R^(B))

In Scheme 9, R and R′ are R^(A) and R^(B) respectively, as defined inStructural Formula (A).

The following compounds were prepared according to Scheme 9.

Br₂ (2.7 mL, 52.0 mmol, 1.2 equiv) was added to a solution of 3 (10.6 g,43.3 mmol) in acetic acid (100 mL) at 25° C. The reaction was stirred at25° C. for 12 h. The resulting mixture was added dropwise to ice-water(400 mL). The mixture was allowed to warm to 25° C. over 1 h. Theresulting suspension was filtered through a pad of Celite. The solid waswashed off with EtOAc. The combined organic layer was dried (Na₂SO₄) andconcentrated to give 14 g of crude 9-1.

Potassium carbonate (8.9 g, 64.5 mmol, 1.5 equiv) and benzyl bromide(11.5 mL, 96.8 mmol, 2.25 equiv) were added to an acetone solution (100mL) of crude 9-1 (14 g, 43 mmol) at 25° C. The reaction was stirred at25° C. for 12 h and concentrated. The resulting mixture was partitionedbetween H₂O and EtOAc. The aqueous layer was extracted with EtOAc. Thecombined EtOAc extracts were dried (Na₂SO₄) and concentrated to yieldcrude 9-2. Flash chromatography on silica gel (100:1 to 30:1hexanes/EtOAc) yielded 15.4 g of compound 9-2 (87% for 2 steps).

Pd(OAc)₂ (227 mg, 1.0 mmol, 0.2 equiv) and P(O-Tol)₃ (462 mg, 1.5 mmol,0.3 euiqv) were added to a DMF solution (10 mL) of 9-2 (2.1 g, 5.06mmol). The reaction was purged with N₂ for 5 min. Et₃N (3.5 mL, 25.3mmol, 5 equiv) and allyloxy-t-butyldimethylsilane (2.2 mL, 10.1 mmol, 2equiv) were added to the reaction. The reaction was heated to 88° C. andstirred at 88° C. for 5 h. The reaction was allowed to cool to 25° C.and quenched with H₂O. The resulting mixture was extracted with EtOAc.The combined EtOAc extracts were dried (Na₂SO₄) and concentrated to givecrude 9-3. Flash chromatography on silica gel (100:0 to 100:1hexanes/EtOAc) yielded 1.2 g of compound 9-3 (47%).

n-BuLi (1.3 mL, 2.07 mmol, 5.5 equiv) was added to a THF solution (5 mL)of diisopropylamine (0.3 mL, 2.07 mmol, 5.5 equiv) at 0° C. The reactionwas stirred at 0° C. for 30 min and cooled to −78° C. TMEDA (0.8 mL,5.64 mmol, 15 equiv) was added to the mixture. To the resulting solutionwas added a THF solution (5 mL) of 9-3 (475 mg, 0.94 mmol, 2.5 equiv).The reaction was stirred at −78° C. for 10 min. A THF solution (5 mL) ofenone (181 mg, 0.376 mmol) was added to the reaction at −78° C. Thereaction was stirred at −78° C. for 30 min and allowed to warm to 25° C.over 1 h, quenched with saturated NH₄Cl, and extracted with EtOAc. Thecombined EtOAc extracts were dried (Na₂SO₄) and concentrated to yieldthe crude product. Preparative reverse phase HPLC purification on aWaters Autopurification system using a Sunfire Prep C18 OBD column [5μm, 19×50 mm; flow rate, 20 mL/min; Solvent A: H₂O with 0.1% HCO₂H;Solvent B: CH₃CN with 0.1% HCO₂H; injection volume: 4.0 mL (CH₃CN);gradient: 100→100% B over 15 min; mass-directed fraction collection].Fractions with the desired MW were collected and concentrated on aRotaVap at 25° C. to remove most of the acetonitrile. The resultingmostly aqueous solution was extracted with EtOAc. The combined EtOAcextracts were dried (Na₂SO₄) and concentrated to give 200 mg of 9-4(59%).

TFA (0.5 mL) was added to a THF/H₂O (2 mL/0.5 mL) solution of 9-4 at 25°C. The reaction was stirred at 25° C. for 1 h. The reaction was quenchedwith sat. NaHCO₃ solution. The reaction was extracted with EtOAc. Thecombined EtOAc extracts were dried (Na₂SO₄) and concentrated to givecrude 9-5. Preparative reverse phase HPLC purification on a WatersAutopurification system using a Sunfire Prep C18 OBD column [5 μm, 19×50mm; flow rate, 20 mL/min; Solvent A: H₂O with 0.1% HCO₂H; Solvent B:CH₃CN with 0.1% HCO₂H; injection volume: 4.0 mL (CH₃CN); gradient:80→100% B over 15 min; mass-directed fraction collection]. Fractionswith the desired MW were collected and concentrated on a RotaVap at 25°C. to remove most of the acetonitrile. The resulting mostly aqueoussolution was extracted with EtOAc. The combined EtOAc extracts weredried (Na₂SO₄) and concentrated to give 80 mg of 9-5 (46%).

Dess-Martin periodinane (18 mg, 0.043 mmol, 1.2 equiv) was added to aCH₂Cl₂ solution (1 mL) of 9-5 (28 mg, 0.036 mmol) at 25° C. The reactionwas stirred at 25° C. for 30 min and diluted with H₂O. The resultingmixture was extracted with CH₂Cl₂. The combined CH₂Cl₂ extracts weredried (Na₂SO₄) and concentrated to give crude 9-6.

Pyrrolidine (15 μL, 0.18 mmol, 5 equiv) was added to a dichloroethanesolution (1 mL) of crude 9-6 (0.036 mmol) at 25° C. The reaction wasstirred at 25° C. for 10 min. HOAc (15 μL) and NaBH(OAc)₃ (15 mg, 0.072mmol, 2 equiv) were added to the reaction. The reaction mixture wasstirred at 25° C. for 1 h and quenched with H₂O. The resulting mixturewas extracted with CH₂Cl₂. The combined CH₂Cl₂ extracts were dried(Na₂SO₄) and concentrated to give crude 9-7-1. Preparative reverse phaseHPLC purification on a Waters Autopurification system using a SunfirePrep C18 OBD column [5 μm, 19×50 mm; flow rate, 20 mL/min; Solvent A:H₂O with 0.1% HCO₂H; Solvent B: CH₃CN with 0.1% HCO₂H; injection volume:4.0 mL (CH₃CN); gradient: 0→100% B over 15 min; mass-directed fractioncollection]. Fractions with the desired MW were collected andconcentrated on a RotaVap at 25° C. to remove most of the acetonitrile.The resulting mostly aqueous solution was extracted with EtOAc. Thecombined EtOAc extracts were dried (Na₂SO₄) and concentrated to give 6mg of 9-7-1 (20% for 2 steps).

Aqueous HF (0.3 mL, 48%) was added to a CH₃CN solution (2 mL) of 9-7-1(6 mg, 0.007 mmol) in a polypropylene tube at 25° C. The reaction wasstirred at 25° C. for 18 h. The resulting mixture was poured into anaqueous solution of K₂HPO₄ (2 g, dissolved in 15 mL water). The mixturewas extracted with EtOAc. The combined EtOAc extracts were dried(Na₂SO₄) and concentrated to yield crude desilyl product.

Palladium on carbon (2 mg, 10 wt %) was added to a HCl/MeOH solution(0.5N, 2 mL) of the crude desilyl product. The reaction was purged withhydrogen and stirred under H₂ (balloon) at 25° C. for 4 h. The reactionmixture was filtered through a small Celite plug. The filtrate wasconcentrated to yield the crude product. Preparative reverse phase HPLCpurification on a Waters Autopurification system using a PhenomenexPolymerx 10μ RP-1 100 A column [10 μm, 150×21.20 mm; flow rate, 20mL/min; Solvent A: 0.05 N HCl/water; Solvent B: CH₃CN; injection volume:4.0 mL (0.05 N HCl/water); gradient: 0→50% B over 7 min, 50→100% over 3min, and 100% over 5 min; mass-directed fraction collection]. Fractionswith the desired MW, eluting at 6.4-8.2 min, were collected andfreeze-dried to yield 1.5 mg of compound 124: ¹H NMR (400 MHz, CD₃OD) δ7.28 (d, J=9.7 Hz, 1H), 4.08 (s, 1H), 3.71-3.63 (m, 2H), 3.32-2.95 (m,7H), 3.04 (s, 3H), 2.96 (s, 3H), 2.81-2.73 (m, 2H), 2.32-1.98 (m, 8H),1.70-1.59 (m, 1H); MS (ESI) m/z 544.18 (M+H).

Compounds 125-127 were prepared similarly to compound 124 using thecorresponding amines in the reductive amination step.

¹H NMR (400 MHz, CD₃OD) δ 7.28 (d, J=9.7 Hz, 1H), 4.08 (s, 1H),3.25-2.94 (m, 5H), 3.04 (s, 3H), 2.96 (s, 3H), 2.89 (s, 6H), 2.80-2.70(m, 2H), 2.32-2.18 (m, 2H), 2.10-2.00 (m, 2H), 1.70-1.58 (m, 1H); MS(ESI) m/z 518.26 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 7.27 (d, J=9.6 Hz, 1H), 4.08 (s, 1H),3.20-2.93 (m, 5H), 3.04 (s, 3H), 2.96 (s, 3H), 2.82-2.72 (m, 2H),2.33-2.19 (m, 2H), 2.04-1.94 (m, 2H), 1.70-1.58 (m, 2H), 1.37 (s, 9H);MS (ESI) m/z 546.20 (M+H).

¹H NMR (400 MHz, CD₃OD) δ 7.28 (d, J=9.7 Hz, 1H), 4.09 (s, 1H), 4.04 (q,J=9.0 Hz, 2H), 3.25-2.95 (m, 5H), 3.04 (s, 3H), 2.97 (s, 3H), 2.84-2.75(m, 2H), 2.32-2.20 (m, 2H), 2.13-2.03 (m, 2H), 1.70-1.58 (m, 1H); MS(ESI) m/z 572.22 (M+H).

Compound 128 was prepared from compound 9-5 by HF treatment following byhydrogenation under similar conditions: ¹H NMR (400 MHz, CD₃OD) δ 7.21(d, J=9.87 Hz, 1H), 4.07 (s, 1H), 3.63-3.57 (m, 2H), 3.20-2.90 (m, 5H),3.04 (s, 3H), 2.96 (s, 3H), 2.75-2.68 (m, 2H), 2.32-2.17 (m, 2H),1.89-1.79 (m, 2H), 1.70-1.57 (m, 1H), 1.25 (d, J=7.2 Hz, 1H); MS (ESI)m/z 491.18 (M+H).

Example 10. Synthesis of Compound 129

The following compounds were prepared according to Scheme 10.

iPrMgCl.LiCl (0.68 mL, 1.2 M, 0.82 mmol, 2 equiv) was added to a THFsolution (5 mL) of 9-2 (170 mg, 0.41 mmol) at 0° C. The reaction wasstirred at 0° C. for 30 min. Mel (0.2 mL, 1.64 mmol, 4 equiv) was addedto the reaction mixture. The reaction was stirred at 0° C. for 30 minand allowed to warm to 25° C. over 1 h. The reaction was quenched withNH₄Cl solution and extracted with EtOAc. The combined EtOAc extractswere dried (Na₂SO₄) and concentrated to give crude 10-1. Flashchromatography on silica gel (30:1 hexanes/EtOAc) yielded 31 mg ofcompound 10-1 (22%).

A THF solution (1 mL) of 10-1 (31 mg, 0.088 mmol, 1.7 equiv) was add toa THF solution (1 mL) of LDA (0.13 mL, 1.3 M, 0.176 mmol, 3.3 equiv) andTMEDA (39 μL, 0.26 mmol, 4.9 equiv). The reaction was stirred at −78° C.for 10 min. A THF solution (1 mL) of enone (26 mg, 0.053 mmol) was addedto the reaction at −78° C. The reaction was stirred at −78° C. for 30min and allowed to warm to 25° C. over 1 h, quenched with saturatedNH₄Cl solution, and extracted with EtOAc. The combined EtOAc extractswere dried (Na₂SO₄) and concentrated to yield the crude 10-2.Preparative reverse phase HPLC purification on a Waters Autopurificationsystem using a Sunfire Prep C18 OBD column [5 μm, 19×50 mm; flow rate,20 mL/min; Solvent A: H₂O with 0.1% HCO₂H; Solvent B: CH₃CN with 0.1%HCO₂H; injection volume: 4.0 mL (CH₃CN); gradient: 80→100% B over 15min; mass-directed fraction collection]. Fractions with the desired MWwere collected and concentrated on a RotaVap at 25° C. to remove most ofthe acetonitrile. The resulting mostly aqueous solution was extractedwith EtOAc. The combined EtOAc extracts were dried (Na₂SO₄) andconcentrated to give 10 mg of 10-2 (26%).

Aqueous HF (0.3 mL, 48%) was added to a CH₃CN solution (2 mL) of 10-2 (6mg, 0.008 mmol) in a polypropylene tube at 25° C. The reaction wasstirred at 25° C. for 18 h. The resulting mixture was poured into anaqueous solution of K₂HPO₄ (2 g, dissolved in 15 mL water). The mixturewas extracted with EtOAc. The combined EtOAc extracts were dried(Na₂SO₄) and concentrated to yield crude 10-3.

Palladium on carbon (2 mg, 10 wt %) was added to a HCl/MeOH solution(0.5N, 2 mL) of the crude 10-3. The reaction was purged with hydrogenand stirred under H₂ (balloon) at 25° C. for 4 h. The reaction mixturewas filtered through a small Celite plug. The filtrate was concentratedto yield the crude product. Preparative reverse phase HPLC purificationon a Waters Autopurification system using a Phenomenex Polymerx 10μ RP-1100 A column [10 μm, 150×21.20 mm; flow rate, 20 mL/min; Solvent A: 0.05N HCl/water; Solvent B: CH₃CN; injection volume: 4.0 mL (0.05 NHCl/water); gradient: 0→70% B over 7 min, 70→100% over 3 min, and 100%over 5 min; mass-directed fraction collection]. Fractions with thedesired MW were collected and freeze-dried to yield 1.5 mg of compound129: ¹H NMR (400 MHz, CD₃OD) δ 7.19 (d, J=9.7 Hz, 1H), 4.07 (s, 1H),3.20-2.93 (m, 3H), 3.03 (s, 3H), 2.96 (s, 3H), 2.31-2.17 (m, 2H), 2.22(s, 3H), 1.69-1.58 (m, 1H); MS (ESI) m/z 447.23 (M+H).

Example 11. Synthesis of Compounds 130-132

In Scheme 11, R⁶, R⁹, and R^(9′) is X, R^(B), and R^(A), respectively,as defined in Structural Formula (A).

Compounds 130-132 were prepared according to Scheme 11.

Example 12. Synthesis of Compounds 133-135

In Scheme 12, R⁹ and R^(9′) is R^(B), and R^(A), respectively, asdefined in Structural Formula (A).

Compounds 133-135 were prepared according to Scheme 12.

Example 13

The antibacterial activities for the compounds of the invention werestudied according to the following protocols.

Minimum Inhibitory Concentration Assay

Frozen bacterial strains were thawed and subcultured onto Mueller HintonBroth (MHB) or other appropriate media (Streptococcus requires blood andHaemophilus requires hemin and NAD). Following incubation overnight, thestrains were subcultured onto Mueller Hinton Agar and again incubatedovernight. Colonies were observed for appropriate colony morphology andlack of contamination. Isolated colonies were selected to prepare astarting inoculum equivalent to a 0.5 McFarland standard. The startinginoculum was diluted 1:125 using MHB for further use. Test compoundswere prepared by dilution in sterile water to a final concentration of5.128 mg/mL. Antibiotics (stored frozen, thawed and used within 3 hoursof thawing) and compounds were further diluted to the desired workingconcentrations.

The assays were run as follows. Fifty μL of MHB was added to wells 2-12of a 96-well plate. One hundred μL of appropriately diluted antibioticswas added to well 1. Fifty μL of antibiotics was removed from well 1 andadded to well 2 and the contents of well 2 mixed by pipetting up anddown five times. Fifty μL of the mixture in well 2 was removed and addedto well 3 and mixed as above. Serial dilutions were continued in thesame manner through well 12. Fifty μL was removed from well 12 so thatall contained 50 μL. Fifty μL of the working inoculum was then added toall test wells. A growth control well was prepared by adding 50 μL ofworking inoculum and 50 μL of MHB to an empty well. The plates were thenincubated at 37° C. overnight, removed from the incubator and each wellwas read on a plate reading mirror. The lowest concentration (MIC) oftest compound that inhibited the growth of the bacteria was recorded.

Example

1 2 3 4 5 6 7 8 9 10 11 12 [Abt] 32 16 8 4 2 1 0.5 0.25 0.125 0.06 0.030.015 grow − − − − − + + + + + + + [Abt] = antibiotic concentration inthe well grow = bacterial growth (cloudiness)

Interpretation: MIC=2 μg/mL

Protocol for Determining Inoculum Concentration (Viable Count)

Ninety μL of sterile 0.9% NaCl was pipetted into wells 2-6 of a 96-wellmicrotiter plate. Fifty 50 μl of the inoculum was pipetted into well 1.Ten μL was removed from well 1 and added it to well 2 followed bymixing. Ten μL was removed from well two and mixed with the contents ofwell 3 and so on creating serial dilutions through well 6. Ten μL wasremoved from each well and spotted onto an appropriate agar plate. Theplate was placed into a CO₂ incubator overnight. The colonies in spotsthat contain distinct colonies were counted. Viable count was calculatedby multiplying the number of colonies by the dilution factor.

Spot from Well 1 2 3 4 5 6 Dilution Factor 10² 10³ 10⁴ 10⁵ 10⁶ 10⁷

Bacterial Strains

Fifteen bacterial strains, listed below, were examined in minimuminhibitory concentration (MIC) assays.

ID Organism Source Resistance Comments Gram Rx SA100 S. aureus ATCC13709 MSSA Smith strain positive (animal model) SA101 S. aureus ATCC29213 MSSA control positive SA158 S. aureus MR, SK75 tet resistant: tetKpositive (efflux) SA161 S. aureus Micromyx, LLC tet resistant: tet(M)positive ribosomal protection EF103 E. faecalis ATCC 29212 tetintermediate/resistant - control positive mechanism not specified EF159E. faecalis MR, DS160 tet resistant: tetM cip-R, ery-I positive (ribprotect) SP106 S. pneumoniae ATCC 49619 wt control positive SP160 S.pneumoniae MR, 54 tet resistant: tet M pen-R, ery-R positive (ribprotect) EC107 E. coli ATCC 25922 wt control negative EC155 E. coli MR,10 tet resistant: tetA negative (efflux) KP109 K. pneumoniae ATCC 13883wt negative KP153 K. pneumoniae MR, 1 tet resistant: tetA cip-R, gen-Rnegative (efflux) EC108 E. cloacae ATCC 13047 wt negative AB110 A.baumanii ATCC 19606 wt negative PA111 P. aeruginosa ATCC 27853 wtcontrol negative MSSA = methicillin susceptible S. aureus wt = wild typeATCC = American Type Culture Collection MR = Marilyn Roberts, Universityof Washington tet = tetracycline cip = ciprofloxacin R = resistant gen =gentamicin ery = erythromycin pen = penicillin

Results

Values of minimum inhibition concentration (MIC) for the compounds ofthe invention are provided in Tables 1a, 1b, 2a, 2b and 3.

TABLE 1a Compound MICs (ug/mL) ID SA101 SA100 SA161 SA158 EF103 EF159SP106 SP160 11 0.125 0.25 0.25 0.0625 0.0625 0.125 ≦0.0156 ≦0.0156 12≦0.0156 ≦0.0156 0.125 0.5 ≦0.0156 0.0625 ≦0.0156 ≦0.0156 13 ≦0.0156≦0.0156 0.0625 0.25 ≦0.0156 0.0625 ≦0.0156 ≦0.0156 14 0.5 0.25 0.5 0.250.125 0.25 0.03 0.125 15 0.5 0.5 0.5 0.25 0.5 0.5 0.016 0.016 16 0.1250.25 0.25 0.5 0.0625 0.0625 ≦0.0156 ≦0.0156 17 0.25 0.5 1 2 0.25 10.0625 0.0625 18 0.0625 ≦0.0156 0.0625 0.125 ≦0.0156 0.0313 ≦0.0156≦0.0156 19 0.125 0.25 0.25 0.25 ≦0.0156 0.125 ≦0.0156 ≦0.0156 20 ≦0.01560.25 0.25 0.5 ≦0.0156 0.0625 ≦0.0156 ≦0.0156 21 0.25 0.25 0.5 4 0.125 1≦0.0156 0.0625 22 1 1 2 4 2 4 0.5 1 23 ≦0.0156 0.5 0.125 0.25 ≦0.01560.0313 ≦0.0156 ≦0.0156 24 0.25 0.25 0.125 0.125 ≦0.0156 ≦0.0156 ≦0.0156≦0.0156 25 0.125 0.125 0.125 0.0313 ≦0.0156 0.0625 ≦0.0156 ≦0.0156 26 11 1 1 0.25 0.5 ≦0.0156 ≦0.0156 27 2 2 4 16 2 4 0.125 0.5 28 1 2 2 1 1 10.125 0.0625 29 4 4 4 2 2 2 1 1 30 2 1 4 2 2 2 2 4 31 1 2 2 1 0.5 1 0.250.25

TABLE 1b MICs (ug/mL) Compound ID EC107 EC155 AB110 PA111 EC108 KP109KP153 11 0.25 2 0.5 16 1 1 2 12 0.25 8 1 16 1 1 4 13 0.125 4 0.25 16 10.5 2 14 0.5 4 0.25 16 2 1 2 15 1 4 0.125 16 4 2 4 16 0.5 8 1 16 1 1 417 2 32 0.5 32 8 4 32 18 0.125 4 0.25 16 0.5 0.5 4 19 0.25 4 0.25 16 2 14 20 0.25 8 1 8 1 1 4 21 1 16 2 16 4 2 8 22 16 >32 2 >32 >32 32 >32 230.125 8 0.125 8 1 0.5 8 24 0.5 8 0.0313 16 2 1 8 25 0.5 8 0.125 32 2 2 826 0.5 4 0.25 16 2 2 4 27 4 32 16 >32 16 8 32 28 2 16 0.5 >32 8 4 16 298 8 8 >32 8 8 8 30 >32 >32 8 >32 >32 >32 >32 31 2 8 0.5 >32 8 4 8

TABLE 2a Compound MICs (ug/mL) ID SA101 SA100 SA161 SA158 EF103 EF159SP106 SP160 32 0.125 0.5 0.25 0.5 0.0625 0.125 ≦0.0156 ≦0.0156 33 0.250.5 1 2 0.25 1 ≦0.0156 0.125 34 ≦0.0156 0.0625 0.0625 0.125 ≦0.0156≦0.0156 ≦0.0156 ≦0.0156 35 0.25 0.25 0.5 0.5 0.125 0.25 0.016 0.016 360.25 0.5 0.5 1 0.25 0.25 ≦0.0156 ≦0.0156 37 8 8 >32 >32 16 16 2 4 38 8 816 32 8 16 2 8 39 2 2 2 16 2 2 0.25 0.5 40 1 1 1 16 1 1 0.0625 0.5 41 11 2 1 1 2 0.125 0.125 42 0.5 1 2 1 0.5 1 0.125 0.0625 43 0.5 0.5 1 0.50.25 0.5 ≦0.0156 ≦0.0156 44 4 4 8 8 8 8 0.5 1 45 0.5 0.5 1 0.5 0.25 0.50.125 0.06 46 0.25 0.25 0.5 0.25 0.25 0.5 0.125 0.125

TABLE 2b MICs (ug/mL) Compound ID EC107 EC155 AB110 PA111 EC108 KP109KP153 32 0.25 8 2 8 1 1 4 33 2 >32 4 >32 16 4 >32 34 0.25 2 0.125 16 10.5 2 35 1 16 0.25 >32 8 4 8 36 2 32 0.125 >32 4 4 3237 >32 >32 >32 >32 >32 >32 >32 38 >32 >32 >32 >32 >32 >32 >32 39 4 >3232 >32 16 16 >32 40 4 >32 8 32 4 8 32 41 4 16 0.5 >32 16 8 16 42 4 160.5 >32 16 8 16 43 1 4 0.125 32 4 2 4 44 32 >32 8 >32 >32 >32 >32 45 1 20.06 >32 4 2 4 46 2 4 0.125 >32 32 4 32

TABLE 3 MIC Values for Compounds of the Invention Compared toSancycline, Minocycline and Tigecycline. SA161 SA101 SA100 MRSA, SA158EF103 EF159 SP106 SP160 Cmpd 29213 13709 tetM tetK 29212 tetM 49619 tetM11 B B B B B B A A 12 A A A B A B A A 13 A A A B A B A A 14 B B B B B BB B 15 B B B B B B A A 16 B B B B B B A A 17 B B B B B B B B 18 B A A BA A A A 19 B B B B A B A A 20 A B B B A B A A 21 B B B B B B A B 22 C BB B B B C B 23 A B A B A A A A 24 B B A B A A A A 25 B B A B A B A A 26C B B B B B A A 27 C C B C B B B B 28 C C B B B B B B 29 C C B B B B C B30 C B B B B B C B 31 C C B B B B B B 32 B B B B B B A A 33 B B B B B BA B 34 A B A B A A A A 35 B B B B B B A A 36 B B B B B B A A 37 C C C CC B C B 38 C C C C B B C B 39 C C B C B B B B 40 C B B C B B B B 41 C BB B B B B B 42 B B B B B B B B 43 B B B B B B A A 44 C C B C B B C B 45B B B B B B B B 46 B B B B B B B B 47 B B B B B B A A 48 B B B B A B A A49 B B B B B B A B 50 B B B B B B B B 51 A A B B A A A A 52 B B B B B BA A 53 C C C C B C B B 54 B B B B B B C B 55 B B B B B B A B 56 B B B BB B B B 57 B B B B B B C B 58 C B B B B B C B 59 B B B B B B B B 60 B BB B B B B B 61 C C C C B B B B 62 C B B C B B B B 63 B B B C B B A B 64B B B B B B A A 65 C C B B B B B B 66 B B B B B B A A 67 C C B B B B B B68 B B B B B B B B 69 C C B B B B C B 70 C C B B B B B B 71 B B B B B BB B 72 B B B B B B A A 73 C B B B B B C B 74 C C C C C B C B 75 B B B CB B B B 76 B B B B B B C C 77 B B B B B B C C 78 B B B B B B C C 79 B BB B B B C C 80 C C B C B B C B 81 B B A B B B C C 82 B B A B B B C B 83B B A B B B C C 84 B B B B B B B B 85 B B B B B B A B 86 B B B B B B B B87 C B B B B B C C 88 B B A B B B A B 89 A A A A A A A A 90 B B A B B BC B 91 B B B B B B C B 92 B B B B B B C B 93 B B B B B B C B 94 B B A BB B C B 95 B B B B B B C B 96 B B B B B B C B 97 B B B B B B C C 98 B BB B B B C B 99 C B B B B B C C 100 B B B B B B C C 101 B B B B B B B B102 C B B B B B B B 103 C C C C B C C B 104 B B B B B B B B 105 C B B CB B B B 106 C B B B B B C B 107 NT NT NT NT NT NT NT NT 108 C C C C C CC C 109 C C C C C C C C 110 C C C C C C C C 111 C C C C C C C C 112 C CC C C C C C 113 C C C C B C B B 114 C C B B B B B B 115 C C B B B B C B116 C C C B B B B B 117 C C C B B B B B 118 C C B B B B B B 119 C C B BB B C B 120 C C C C C C C C 121 C C C C B C C B 122 C C C C C C C B 123C C C C C C C C 124 C C C C B C C B 125 C C C B B B C B 126 C C C C C CC C 127 C B B B B B C B 128 C C B C B B C C 129 C B B B B B C B 131 C CB B C B C C 132 C C C B B B C B 133 B B B B B B B B 130 C B B B B B C B134 B B B B B B A A 135 B B B B B B A A Minocycline 0.06 0.06 8 0.03 116 0.016 2 Sancycline 0.5 1 4 8 8 0.25 8 Tigecycline 0.06 0.06 0.1250.06 0.03 0.06 0.016 0.016 EC107 EC155 AB110 PA111 EC108 KP109 KP153Cmpd 25922 tetA 19606 27853 13047 13883 tetA  11 B B C B B B B  12 B B CB B B B  13 B B B B B B B  14 B B B B B B B  15 B B B B B B B  16 B B CB B B B  17 B B C C B B B  18 B B B B B B B  19 B B B B B B B  20 B B CA B B B  21 B B C B B B B  22 C C C C C C C  23 B B B A B B B  24 B B AB B B B  25 B B B C B B B  26 B B B B B B B  27 B B C C C B B  28 B B CC B B B  29 B B C C B B B  30 C C C C C C C  31 B B C C B B B  32 B B CA B B B  33 B C C C C B C  34 B B B B B B B  35 B B B C B B B  36 B B BC B B B  37 C C C C C C C  38 C C C C C C C  39 B C C C C C C  40 B C CC B B B  41 B B C C C B B  42 B B C C C B B  43 B B B C B B B  44 C C CC C C C  45 B B A C B B B  46 B B B C C B B  47 B B C A B B B  48 A B AA A A A  49 B B B C B B A  50 B B B C B B B  51 B B B B B B B  52 B B BC B B A  53 C C C C C C C  54 C C C C C C C  55 B C C C B B C  56 B C CC C B C  57 C C C C C C C  58 C C C C C C C  59 B C C C C C C  60 B C CC B B B  61 C C C C C C C  62 C C C C C C C  63 B C C C B B C  64 B B CB B B B  65 B B C C B B B  66 B B B C B B B  67 B B C C B B B  68 B B CB B B B  69 B C C C B C B  70 B C C C B B C  71 B C C C C B C  72 B C CC C C C  73 C C C C C C C  74 C C C C C C C  75 B C C C B B C  76 C C CC C C C  77 C C C C C C C  78 C C C C C C C  79 C C C C C C C  80 C C CC C C C  81 C C C C C C C  82 C C C C C C C  83 C C C C C C C  84 C C CC C C C  85 B C C C B B C  86 C C C C C C C  87 C C C C C C C  88 B B CA B B B  89 A B A A A A A  90 C C C C C C C  91 C C C C C C C  92 C C CC C C C  93 C C C C C C C  94 C C C C C C C  95 C C C C C C C  96 C C CC C C C  97 C C C C C C C  98 C C C C C C C  99 C C C C C C C 100 C C CC C C C 101 B B C C B B C 102 B B C C B B B 103 B C C C B C C 104 B C CC B B C 105 B C C C C C C 106 C C C C C C C 107 NT NT NT NT NT NT NT 108C C C C C C C 109 C C C C C C C 110 C C C C C C C 111 C C C C C C C 112C C C C C C C 113 B C C C B B C 114 B B C C B B B 115 B B C C C B B 116B C C C B B B 117 B C C C B B C 118 B C C C B B B 119 C B C C C C B 120C C C C C C C 121 C C C C C C C 122 C C C C C C C 123 C C C C C C C 124C C C C C C C 125 C C C C C C C 126 C C C C C C C 127 C C C C C C C 128C C C C C C C 129 C C C C C C B 131 C B C C C C B 132 C B C C C C B 133B B C C C B B 130 C C C C C C C 134 B B C C B B B 135 B B C C B B BMinocycline 0.5 8 0.06 16 2 1 8 Sancycline 8 32 0.25 >32 8 8 32Tigecycline 0.03 0.5 0.25 8 0.25 0.125 1 A = lower than or equal tolowest MIC among three control compounds; B = greater than lowest MICamong three control compounds, but lower than highest MIC among threecontrol compounds; C = greater than MIC of all three control compounds.The lowest concentration limit of the assay is 0.016. Thus, reportedvalues in the assay of 0.016 represent an MIC of 0.016 or lower.

ED₅₀ IV Study

Compounds of the present invention were evaluated for efficacy in amouse systemic infection model against a susceptible S. aureus isolate,ATCC13709. Mice were infected via intraperitoneal injection with abacterial load that would result in 0% survival within 48 hours afterinfection. Mice received the test compounds 60 minutes post infectionvia intravenous injection. Infection control groups did not receivetreatment. Survival was assessed over a 48 hour period. Percent survivalwas calculated and PD₅₀ values determined using Probit analysis.

Tet-R Sepsis Study

The protocol for tetracycline-resistance sepsis study was similar to theprotocol for EV₅₀ IV study described above except the infection modelused S. aureus SA161, a tetracycline-resistant strain.

GN Sepsis

The protocol for GN sepsis study was similar to the protocol for EV₅₀ IVstudy described above except the infection model used E. coli ATCC25922.

Metabolic Stability

A stock solution of the test compounds were prepared by dissolving thecompound in DMSO to a final concentration of 1.0 mg/mL.

The analytes and internal standard were diluted and infused into theLC/MS system to determine optimal ionization, polarity and MS/MSfragmentation for selection of specific MRM (multiple reactionmonitoring) transitions. Generic chromatographic conditions weredeveloped with a less than 5 minute cycle time.

Pooled human microsome assays were conducted at 0.2 mg/mL protein withan NADPH generating cofactor system (1.3 mM NADP+, 3.3 mMglucose-6-phosphate, 0.4 U/mL glucose-6-phosphate dehydrogenase, and 3.3mM magnesium chloride).

Test compounds were diluted from DMSO stocks to 1 M reactionconcentrations. Time points were removed at 0 and 60 minutes. After aprotein crash with acetonitrile, samples are analyzed by LC/MS/MS. Peakareas are compared to the time 0 sample, and the percent analyteremaining is calculated. A reaction containing no cofactor was used tocontrol for analyte loss due to non-specific binding, thermaldegradation and solubility.

SA MIC90

Twenty randomly selected clinical isolates of S. aureus were used todetermine the minimal inhibitory concentration (MIC) of test compoundsfor 90% of the isolates (MIC₉₀). MICs were performed by microtiter brothdilution in a 96-well format according to Clinical Laboratory StandardsInstitute (CLSI) guidelines, as described above.

Viable counts were determined by 10-fold serial dilution. Dilutions wereprepared in sterile 0.9% NaCl. Ten microliters of the inoculum and ofeach of 5 dilutions were plated onto blood or Mueller Hinton agarplates, incubated overnight at 37° C. with 5% CO₂, and counted.

TetR MIC90

Ten isolates selected based on resistance to tetracycline were used todetermine the MIC₉₀ as described above.

EC MIC90

Twenty randomly selected clinical isolates of E. coli were used todetermine the MIC₉₀ as described above.

Protein Binding

Test compounds were prepared as 1.0 mg/mL stock solutions in DMSO. Theanalytes and internal standard were diluted and infused into the LC/MSsystem to determine optimal ionization, polarity and MS/MS fragmentationfor selection of specific MRM (multiple reaction monitoring)transitions. Generic chromatographic conditions were developed with aless than 5 minute cycle time.

The DMSO stocks were diluted to 1 and 10 μg/mL in human plasma andincubated in RED devices for 4 hours at 37° C. The time point wasremoved at the end of the incubation period. After a protein crash withacetonitrile, the samples were analyzed by LC/MS/MS. Peak areas for thebuffer (receiver) and sample (donor) chambers were compared and theprotein bound fraction is calculated. Analysis was conducted induplicate.

Thigh Burden

Female CD-1 mice were pre-treated with cyclophosphamide to render themice neutropenic. Mice were infected with S. aureus ATCC13709 viainjection into the right thigh muscle of 0.1 ml per mouse. One and ahalf hours post infection mice were treated IV with test compounds indoses ranging from 0.3 to 30 mg/kg or 0.3 to 20 mg/kg. Four mice weretreated with each drug concentration. Twenty-four hours post treatment,mice were euthanized by CO₂ inhalation. The right thighs of the micewere aseptically removed, weighed, homogenized, serially diluted, andplated on TSA medium. The plates were incubated overnight at 37° C. in5% CO₂. Colony forming units per gram of thigh was calculated byenumerating the plated colonies then adjusting for serial dilutions andthe weight of the thigh.

The results for the biological activity studies described above arelisted in Table 4.

TABLE 4 Meta SA TetR EC Prot TetR GN Thigh ED50 Stab MIC MIC MIC Bindsep- sep- Bur- IV % 90 90 90 % sis sis den

0.3 104% 0.03 2 0.5 90 0.4 6.2 4.41 log10 de- crease

0.55  72% 0.03 4 1 95.6 3.5 14.3 5.01 log10 de- crease

0.07  85% 0.03 4 0.5 87.1

0.3  86% 0.03 2 0.5 79.4 1 4.36 5.24 log10 de- crease

0.55  65% 0.03 8 2 87.9

0.3  96% 0.06 2 1 96.9

0.073 109% 0.06 4 4 93.6 0.62 4.3 4.12 log10 de- crease

0.97  55% 0.06 1 0.5 69.1 0.55 2.1 4.89 log10 de- crease ED₅₀: mg/kgrequired to protect 50% of mice in septicemia model with S. aureus Smithstrain. Meta Stab %: % compound remaining after 60 min, 37° C.incubation with human liver microsomes. SA MIC90: concentration (ug/mL)required to inhibit 90% of S. aureus clinical isolates (n = 20). TetRMIC90: concentration (ug/mL) required to inhibit 90% oftetracycline-resistant S. aureus (n = 10). EC MIC90: concentration(ug/mL) required to inhibit 90% of E. coli clinical isolates (n = 20).Prot Bind: % Protein binding in human plasma with compound at 10 uM.TetR sepsis: mg/kg required to protect 50% of mice in septicemia modelwith tetracycline-resistant S. aureus strain. GN sepsis: mg/kg requiredto protect 50% of mice in septicemia model with E. coli strain. ThighBurden: maximum log10 reduction in bacteria compared to untreatedcontrol. Maximum log10 reduction: maximum bacterial count (log10)reduction achieved at tested doses.

While this invention has been particularly shown and described withreferences to example embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims.

1. A compound represented by Structural Formula (A):

or a pharmaceutically acceptable salt thereof, wherein: X is selectedfrom hydrogen, —(C₁-C₇)alkyl, carbocyclyl, aryl and heteroaryl; Y isselected from hydrogen, —(C₁-C₇)alkyl, carbocyclyl,—(C₁-C₄)alkylene-N(R^(A))(R^(B)),—(C₁-C₄)alkylene-N(R^(F))—C(O)—[C(R^(D))(R^(E))]₀₋₄—N(R^(A))(R^(B)),—CH═N—OR^(A), —N(R^(A))(R^(B)),—N(R^(F))—C(O)—[C(R^(D))(R^(E))]₁₋₄—N(R^(A))(R^(B)), —N(R^(F))—C(O)—N(R^(A))(R^(B)), —N(R^(F))—C(O)—(C₁-C₆)alkyl,—N(R^(F))—C(O)-heterocyclyl, —N(R^(F))—C(O)-heteroaryl,—N(R^(F))—C(O)-carbocyclyl, —N(R^(F))—C(O)-aryl,—N(R^(F))—S(O)_(m)—(C₁-C₄)alkylene-N(R^(A)) (R^(B)),—N(R^(F))—S(O)_(m)—(C₁-C₄)alkylene-carbocyclyl, and—N(R^(F))—S(O)_(m)—(C₁-C₄)alkylene-aryl wherein: at least one of X and Yis not hydrogen; each R^(A) and R^(B) are independently selected fromhydrogen, (C₁-C₇)alkyl, —O—(C₁-C₇)alkyl, —(C₀-C₆)alkylene-carbocyclyl,—(C₀-C₆)alkylene-aryl, —(C₀-C₆)alkylene-heterocyclyl,—(C₀-C₆)alkylene-heteroaryl, —(C₁-C₆)alkylene-O-carbocyclyl,—(C₁-C₆)alkylene-O-aryl, —(C₁-C₆)alkylene-O-heterocyclyl,—(C₁-C₆)alkylene-O-heteroaryl, —S(O)_(m)—(C₁-C₆)alkyl,—(C₀-C₄)alkylene-S(O)_(m)-carbocyclyl, —(C₀-C₄)alkylene-S(O)_(m)-aryl,—(C₀-C₄)alkylene-S(O)_(m)-heterocyclyl and—(C₀-C₄)alkylene-S(O)_(m)-heteroaryl; or R^(A) and R^(B) taken togetherwith the nitrogen atom to which they are bound form a heterocyclyl orheteroaryl, wherein the heterocycle or heteroaryl optionally comprises 1to 4 additional heteroatoms independently selected from N, S and O; eachR^(D) and each R^(E) is independently selected from hydrogen,(C₁-C₆)alkyl, carbocyclyl, aryl, heterocyclyl, heteroaryl, or anaturally occurring amino acid side chain moiety, or R^(D) and R^(E)taken together with the carbon atom to which they are bound form a 3-7membered carbocyclyl, or a 4-7 membered heterocyclyl, wherein theheterocyclyl formed by R^(D) and R^(E) optionally comprises one to twoadditional heteroatoms independently selected from N, S and O; R^(F) isselected from hydrogen, (C₁-C₇)alkyl, carbocyclyl, aryl and heteroaryl;and m is 1 or 2, wherein: each carbocyclyl, aryl, heterocyclyl orheteroaryl is optionally and independently substituted with one or moresubstituents independently selected from halo, —(C₁-C₄)alkyl, —OH, ═O,—O—(C₁-C₄)alkyl, —(C₁-C₄)alkyl-O—(C₁-C₄)alkyl, halo-substituted—(C₁-C₄)alkyl, halo-substituted —O—(C₁-C₄)alkyl, —C(O)—(C₁-C₄)alkyl,—C(O)-(fluoro-substituted-(C₁-C₄)alkyl), —S(O)_(m)—(C₁-C₄)alkyl,—N(R^(G))(R^(G)), and CN; each alkyl in the group represented by R^(A),R^(B), R^(D) and R^(E) is optionally and independently substituted withone or more substituents independently selected from halo,—(C₁-C₄)alkyl, —OH, —O—(C₁-C₇)alkyl, —(C₁-C₄)alkyl-O—(C₁-C₄)alkyl,fluoro-substituted-(C₁-C₄)alkyl, —S(O)_(m)—(C₁-C₄)alkyl, and—N(R^(G))(R^(G)), wherein each R^(G) is hydrogen or (C₁-C₄)alkyl,wherein each alkyl in the group represented by R^(G) is optionally andindependently substituted with one or more substituents independentlyselected from —(C₁-C₄)alkyl, (C₃-C₆)cycloalkyl, halo, —OH,—O—(C₁-C₄)alkyl, and (C₁-C₄)alkyl-O—(C₁-C₄)alkyl.
 2. The compound ofclaim 1, wherein the compound is represented by the following StructuralFormula:

or a pharmaceutically acceptable salt thereof, wherein: R¹ and R² areeach independently selected from hydrogen, (C₁-C₇)alkyl,(C₃-C₆)cycloalkyl(C₁-C₄)alkyl, (C₁-C₇)alkoxy(C₁-C₄)alkyl,(C₃-C₆)cycloalkoxy(C₁-C₄)alkyl, (C₃-C₆)cycloalkyl, aryl,aryl(C₁-C₄)alkyl, aryloxy(C₁-C₄)alkyl, arylthio(C₁-C₄)alkyl,arylsufinyl(C₁-C₄)alkyl, arylsulfonyl(C₁-C₄)alkyl, and —O—(C₁-C₇)alkyl,or R¹ and R² taken together with the nitrogen atom to which they arebonded form a monocyclic or bicyclic heteroaryl, or a monocyclic, fusedbicyclic, bridged bicyclic or spiro bicyclic heterocycle, wherein theheteroaryl or heterocycle optionally contains one or two additionalheteroatoms independently selected from N, O and S; and wherein eachalkyl, cycloalkyl, alkoxy and cycloalkoxy moiety in the groupsrepresented by R¹ and R² and each heterocycle represented by NR¹R² takentogether is optionally substituted with one or more substituentsindependently selected from the group consisting of (C₁-C₄)alkyl, halo,—OH, (C₁-C₄)alkoxy, (C₁-C₄)alkylthio, (C₁-C₄)alkylsulfinyl,(C₁-C₄)alkylsulfonyl, (C₁-C₄)alkoxy(C₁-C₄)alkyl, and —N(R³)(R⁴); andeach aryl, aryloxy, arylthio, arylsufinyl and arylsulfonyl moiety in thegroups represented by R¹ and R² and each heteroaryl represented by NR¹R²taken together is optionally substituted with one or more substituentsindependently selected from the group consisting of (C₁-C₄)alkyl, halo,—OH, (C₁-C₄)alkoxy, —S—(C₁-C₄)alkyl, —S(O)(C₁-C₄)alkyl,—S(O)₂(C₁-C₄)alkyl, (C₁-C₄)alkoxy(C₁-C₄)alkyl, —N(R³)(R⁴); —CN,halo(C₁-C₄)alkyl, and halo(C₁-C₄)alkoxy, and R³ and R⁴ are eachindependently selected from the group consisting of —H and (C₁-C₄)alkyl,wherein the (C₁-C₄)alkyl represented by R³ and R⁴ is optionallysubstituted with one or more substituents independently selected fromthe group consisting of (C₁-C₄)alkyl, halo, —OH, (C₁-C₄)alkoxy, and(C₁-C₄)alkoxy(C₁-C₄)alkyl.
 3. The compound of claim 2, wherein thecompound is represented by the following Structural Formula:

or a pharmaceutically acceptable salt thereof, wherein. R¹ and R² areeach independently selected from hydrogen, (C₁-C₇)alkyl,(C₃-C₆)cycloalkyl(C₁-C₄)alkyl, (C₁-C₇)alkoxy(C₁-C₄)alkyl,(C₃-C₆)cycloalkoxy(C₁-C₄)alkyl, (C₃-C₆)cycloalkyl, aryl,aryl(C₁-C₄)alkyl, aryloxy(C₁-C₄)alkyl, arylthio(C₁-C₄)alkyl,arylsufinyl(C₁-C₄)alkyl, arylsulfonyl(C₁-C₄)alkyl; or R¹ and R² takentogether with the nitrogen atom to which they are bonded form amonocyclic or bicyclic heteroaryl, or a monocyclic, fused bicyclic,bridged bicyclic or spiro bicyclic heterocycle, wherein the heteroarylor heterocycle optionally contains one or two additional heteroatomsindependently selected from N, O and S.
 4. The compound of claim 3,wherein R¹ is hydrogen or a (C₁-C₄)alkyl.
 5. The compound of claim 3,wherein R² is selected from (C₁-C₇)alkyl, (C₃-C₆)cycloalkyl(C₁-C₄)alkyl,(C₁-C₇)alkoxy(C₁-C₄)alkyl, phenyl, phenyl(C₁-C₄)alkyl, (C₃-C₆)cycloalkyland halo(C₁-C₄)alkyl, wherein each alkyl, alkoxy and cycloalkyl moietyin the groups represented by R² is optionally substituted with one ormore substituents independently selected from the group consisting of(C₁-C₄)alkyl and halo; and each phenyl moiety in the groups representedby R² is optionally substituted with one or more substituentsindependently selected from the group consisting of (C₁-C₄)alkyl, halo,(C₁-C₄)alkoxy, (C₁-C₄)alkoxy(C₁-C₄)alkyl, —CN, halo(C₁-C₄)alkyl, andhalo(C₁-C₄)alkoxy.
 6. The compound of any one of claim 3, wherein R¹ isselected from hydrogen, methyl and ethyl.
 7. The compound of claim 6,wherein R² is selected from the group consisting of cyclopropyl,cyclobutyl, cyclopentyl, cyclopropylmethyl, cyclobutylmethyl, phenyl,benzyl, —(CH₂)₂—O—CH₃, —(CH₂)₃—OCH₃, —C(CH₃)₃, —CH(CH₃)₂, —CH₂C(CH₃)₃,—CH₂CH(C H₃)₂, —CH₂—CF₃, —(CH₂)₂—CH₂F, and —(CH₂)_(n)CH₃; n is 0, 1, 2,3, 4, 5 or 6; and wherein the phenyl or benzyl group represented by R²is optionally substituted with one or two substituents independentlyselected from the group consisting of (C₁-C₄)alkyl, halogen,(C₁-C₄)alkoxy, (C₁-C₄)alkoxy(C₁-C₄)alkyl, —CN, halo(C₁-C₄)alkyl, andhalo(C₁-C₄)alkoxy.
 8. The compound of claim 7, wherein R² is selectedfrom cyclopropyl, cyclopropylmethyl, cyclobutyl, cyclopentyl,cyclohexyl, —(CH₂)₂—O—CH₃, —C(CH₃)₃, —CH(CH₃)₂, —CH₂—CF₃, —CH₂CH(CH₃)₂,—CH₃ and —CH₂CH₃.
 9. The compound of claim 3, wherein R¹ and R² takentogether with the nitrogen atom to which they are bonded form amonocyclic or bicyclic heteroaryl, or a monocyclic, fused bicyclic,bridged bicyclic or spiro bicyclic heterocycle, wherein the heteroarylor heterocycle optionally contains one additional heteroatom selectedfrom N, O and S; and the heterocycle is optionally substituted with oneor more substituents independently selected from the group consisting of(C₁-C₄)alkyl, halo, —OH, (C₁-C₄)alkoxy, (C₁-C₄)alkylthio,(C₁-C₄)alkylsulfinyl, (C₁-C₄)alkylsulfonyl, (C₁-C₄)alkoxy(C₁-C₄)alkyl,and —N(R³)(R⁴); and the heteroaryl is optionally substituted with one ormore substituents independently selected from the group consisting of(C₁-C₄)alkyl, halo, —OH, (C₁-C₄)alkoxy, —S—(C₁-C₄)alkyl,—S(O)(C₁-C₄)alkyl, —S(O)₂(C₁-C₄)alkyl, (C₁-C₄)alkoxy(C₁-C₄)alkyl,—N(R³)(R⁴), —CN, halo(C₁-C₄)alkyl, and halo(C₁-C₄)alkoxy.
 10. Thecompound of claim 9, wherein R¹ and R² taken together with the nitrogenatom to which they are bonded form a heterocycle selected from the groupconsisting of azetidine, pyrrolidine, morpholine, piperidine,octahydrocyclopenta[c]pyrrol, isoindoline, and azabicyclo[3.1.0]hexane,wherein the heterocycle is optionally substituted with one or moresubstituents independently selected from the group consisting of(C₁-C₄)alkyl, halogen, —OH, (C₁-C₄)alkoxy, —S—(C₁-C₄)alkyl,—S(O)(C₁-C₄)alkyl, —S(O)₂(C₁-C₄)alkyl, (C₁-C₄)alkoxy(C₁-C₄)alkyl, and—N(R³)(R⁴).
 11. The compound of claim 10, wherein the heterocycle isoptionally substituted with halogen, methoxy, hydroxy, methoxymethyl ordimethylamino group.
 12. The compound of claim 3, wherein: a) R¹ ismethyl, and R² is cyclopropyl; b) R¹ is hydrogen, and R² is cyclopropyl;c) R¹ is hydrogen, and R² is cyclobutyl; d) R¹ is methyl, and R² iscyclobutyl; e) R¹ is hydrogen, and R² is cyclopropylmethyl; f) R¹ ishydrogen, and R² is cyclobutylmethyl; g) R¹ is hydrogen, and R² isbenzyl; h) R¹ is hydrogen, and R² is methoxypropyl; i) R¹ is hydrogen,and R² is methoxyethyl; j) R¹ is hydrogen, and R² is phenyl; k) R¹ ismethyl, and R² is t-butyl; l) R¹ is hydrogen, and R² is t-butyl; m) R¹is hydrogen, and R² is methyl; n) R¹ is hydrogen, and R² is ethyl; o) R¹is hydrogen, and R² is propyl; p) R¹ is hydrogen, and R² is butyl; q) R¹is hydrogen, and R² is pentyl; r) R¹ is hydrogen, and R² is hexyl; s) R¹is hydrogen, and R² is heptyl; t) R¹ is methyl, and R² is methyl; u) R¹is hydrogen, and R² is isopropyl; v) R¹ is hydrogen, and R² is2,2-dimethylpropyl; w) R¹ is hydrogen, and R² is trifluoroethyl; x) R¹is hydrogen, and R² is 2-methylpropyl; y) R¹ is hydrogen, and R² is3-fluoropropyl; z) R¹ is ethyl, and R² is ethyl; a1) R¹ is methyl, andR² is methyl; b1) R¹ is hydrogen, and R² is hydrogen; c1) R¹ ishydrogen, and R² is cyclopentyl; d1) R¹ is methyl, and R² iscyclopentyl; or e1) R¹ is methyl, and R² is propyl, or apharmaceutically acceptable salt of any of the foregoing.
 13. Thecompound of claim 3, wherein R¹ and R² taken together with the nitrogenatom to which they are bonded form a group selected from: a)azetidin-1-yl; b) 3-fluoroazetidin-1-yl; c) 3-methylazetidin-1-yl; d)3-methoxyazetidin-1-yl; e) pyrrolidin-1-yl; f) morpholin-4-yl; g)3-fluoropyrrolidin-1-yl; h) 3-hydroxypyrrolidin-1-yl; i)3-N,N-dimethylaminopyrrolidin-1-yl; j) 2-methoxymethylpyrrolidin-1-yl;k) piperidin-1-yl; l) octahydrocyclopenta[c]pyrrol-2-yl; m)isoindolin-2-yl; and n) 3-azabicyclo[3.1.0]hexan-3-yl, or apharmaceutically acceptable salt of any of the foregoing.
 14. Thecompound of claim 3, wherein R¹ is hydrogen or a (C₁-C₄)alkyl; and R² isselected from (C₁-C₇)alkyl, (C₃-C₆)cycloalkyl(C₁-C₄)alkyl,(C₁-C₇)alkoxy(C₁-C₄)alkyl, phenyl, phenyl(C₁-C₄)alkyl, (C₃-C₆)cycloalkyland halo(C₁-C₄)alkyl, wherein each alkyl, alkoxy and cycloalkyl moietyin the groups represented by R² is optionally substituted with one ormore substituents independently selected from the group consisting of(C₁-C₄)alkyl and halo; and each phenyl moiety in the groups representedby R² is optionally substituted with one or more substituentsindependently selected from the group consisting of (C₁-C₄)alkyl, halo,(C₁-C₄)alkoxy, (C₁-C₄)alkoxy(C₁-C₄)alkyl, —CN, halo(C₁-C₄)alkyl, andhalo(C₁-C₄)alkoxy; or R¹ and R² taken together with the nitrogen atom towhich they are bonded form a monocyclic or bicyclic heteroaryl, or amonocyclic, fused bicyclic, bridged bicyclic or spiro bicyclicheterocycle, wherein the heteroaryl or heterocycle optionally containsone additional heteroatom selected from N, O and S; and the heterocycleis optionally substituted with one or more substituents independentlyselected from the group consisting of (C₁-C₄)alkyl, halo, —OH,(C₁-C₄)alkoxy, (C₁-C₄)alkylthio, (C₁-C₄)alkylsulfinyl,(C₁-C₄)alkylsulfonyl, (C₁-C₄)alkoxy(C₁-C₄)alkyl, and —N(R³)(R⁴); and theheteroaryl is optionally substituted with one or more substituentsindependently selected from the group consisting of (C₁-C₄)alkyl, halo,—OH, (C₁-C₄)alkoxy, —S—(C₁-C₄)alkyl, —S(O)(C₁-C₄)alkyl,—S(O)₂(C₁-C₄)alkyl, (C₁-C₄)alkoxy(C₁-C₄)alkyl, —N(R³)(R⁴), —CN,halo(C₁-C₄)alkyl, and halo(C₁-C₄)alkoxy.
 15. The compound of claim 2,wherein R¹ is hydrogen, methyl, ethyl, methoxy or tert-butoxy; R² isselected from (C₁-C₇)alkyl, (C₃-C₆)cycloalkyl(C₁-C₄)alkyl,(C₁-C₇)alkoxy(C₁-C₄)alkyl, phenyl, (C₃-C₆)cycloalkyl, andfluoro(C₁-C₄)alkyl; or R¹ and R² taken together with the nitrogen atomto which they are bonded form a ring selected from pyrrolidinyl,morpholinyl, azetidinyl, piperidinyl, octahydrocyclopenta[c]pyrrolyl,isoindolinyl, indazolyl, imidazolyl, pyrazolyl, triazolyl, andtetrazolyl, wherein the ring formed by R¹ and R² taken together with thenitrogen atom to which they are bonded is optionally substituted withfluoro, —OH, —OCH₃, or N(CH₃)₂.
 16. The compound of claim 3, wherein: R¹hydrogen, methyl, or ethyl R² is selected from methyl, ethyl, n-propyl,isopropyl, n-butyl, 2,2-dimethylpropyl, t-butyl, isobutyl, n-pentyl,(C₄-C₆)cycloalkyl, (C₃-C₅)cycloalkylmethyl, methoxyethyl, and2-fluoroethyl; or R¹ and R² taken together with the nitrogen atom towhich they are bonded form a ring selected from azetidinyl,pyrrolidinyl, piperidinyl, tetrazolyl, oroctahydrocyclopenta[c]pyrrolyl, and wherein the ring formed by R¹ and R²taken together with the nitrogen atom to which they are bonded isoptionally substituted with fluoro.
 17. A pharmaceutical compositioncomprising a pharmaceutically acceptable carrier or diluent and acompound of claim 1 or a salt thereof.
 18. A method for treating orpreventing an infection or colonization in a subject comprisingadministering to the subject an effective amount of a compound of claim1 or a salt thereof.
 19. The method of claim 18, wherein the infectionis caused by a Gram-positive organism.
 20. (canceled)
 21. The method ofclaim 18, wherein the infection is caused by a Gram-negative organism.22-30. (canceled)